Inhibitors of Akt activity

ABSTRACT

The instant invention provides for compounds that inhibit Akt activity. In particular, the compounds disclosed selectively inhibit one or two of the Akt isoforms. The invention also provides for compositions comprising such inhibitory compounds and methods of inhibiting Akt activity by administering the compound to a patient in need of treatment of cancer.

PRIORITY CLAIM

This application is a §371 application of PCT/US06/004715 that was filedon Feb. 10, 2006, which claims priority from the U.S. ProvisionalApplication No. 60/652,737 filed on Feb. 14, 2005, now expired.

BACKGROUND OF THE INVENTION

The present invention relates to compounds which are inhibitors of theactivity of one or more of the isoforms of the serine/threonine kinase,Akt (also known as PKB; hereinafter referred to as “Akt”). The presentinvention also relates to pharmaceutical compositions comprising suchcompounds and methods of using the instant compounds in the treatment ofcancer.

Apoptosis (programmed cell death) plays essential roles in embryonicdevelopment and pathogenesis of various diseases, such as degenerativeneuronal diseases, cardiovascular diseases and cancer. Recent work hasled to the identification of various pro- and anti-apoptotic geneproducts that are involved in the regulation or execution of programmedcell death. Expression of anti-apoptotic genes, such as Bcl2 orBcl-x_(L), inhibits apoptotic cell death induced by various stimuli. Onthe other hand, expression of pro-apoptotic genes, such as Bax or Bad,leads to programmed cell death (Adams et al. Science, 281:1322-1326(1998)). The execution of programmed cell death is mediated by caspase-1related proteinases, including caspase-3, caspase-7, caspase-8 andcaspase-9 etc (Thornberry et al. Science, 281:1312-1316 (1998)).

The phosphatidylinositol 3′-OH kinase (PI3K)/Akt pathway appearsimportant for regulating cell survival/cell death (Kulik et al. Mol.Cell. Biol. 17:1595-1606 (1997); Franke et al, Cell, 88:435-437 (1997);Kauffmann-Zeh et al. Nature 385:544-548 (1997) Hemmings Science,275:628-630 (1997); Dudek et al., Science, 275:661-665 (1997)). Survivalfactors, such as platelet derived growth factor (PDGF), nerve growthfactor (NGF) and insulin-like growth factor-1 (IGF-1), promote cellsurvival under various conditions by inducing the activity of PI3K(Kulik et al. 1997, Hemmings 1997). Activated PI3K leads to theproduction of phosphatidylinositol(3,4,5)-triphosphate(PtdIns(3,4,5)-P3), which in turn binds to, and promotes the activationof, the serine/threonine kinase Akt, which contains a pleckstrinhomology (PH)-domain (Franke et al Cell, 81:727-736 (1995); HemmingsScience, 277:534 (1997); Downward, Curr. Opin. Cell Biol. 10:262-267(1998), Alessi et al., EMBO J. 15: 6541-6551 (1996)). Specificinhibitors of PI3K or dominant negative Akt mutants abolishsurvival-promoting activities of these growth factors or cytokines. Ithas been previously disclosed that inhibitors of PI3K (LY294002 orwortmannin) blocked the activation of Akt by upstream kinases. Inaddition, introduction of constitutively active PI3K or Akt mutantspromotes cell survival under conditions in which cells normally undergoapoptotic cell death (Kulik et al. 1997, Dudek et al. 1997).

Three members of the Akt subfamily of second-messenger regulatedserine/threonine protein kinases have been identified and termedAkt1/PKBα, Akt2/PKBβ, and Akt3/PKBγ (hereinafter referred to as “Akt1”,“Akt2” and “Akt3”), respectively. The isoforms are homologous,particularly in regions encoding the catalytic domains. Akts areactivated by phosphorylation events occurring in response to PI3Ksignaling. PI3K phosphorylates membrane inositol phospholipids,generating the second messengers phosphatidyl-inositol3,4,5-trisphosphate and phosphatidylinositol 3,4-bisphosphate, whichhave been shown to bind to the PH domain of Akt. The current model ofAkt activation proposes recruitment of the enzyme to the membrane by3′-phosphorylated phosphoinositides, where phosphorylation of theregulatory sites of Akt by the upstream kinases occurs (B. A. Hemmings,Science 275:628-630 (1997); B. A. Hemmings, Science 276:534 (1997); J.Downward, Science 279:673-674 (1998)).

Phosphorylation of Akt1 occurs on two regulatory sites, Thr³⁰⁸ in thecatalytic domain activation loop and on Ser⁴⁷³ near the carboxy terminus(D. R. Alessi et al. EMBO J. 15:6541-6551 (1996) and R. Meier et al. J.Biol. Chem. 272:30491-30497 (1997)). Equivalent regulatoryphosphorylation sites occur in Akt2 and Akt3. The upstream kinase, whichphosphorylates Akt at the activation loop site has been cloned andtermed 3′-phosphoinositide dependent protein kinase 1 (PDK1). PDK1phosphorylates not only Akt, but also p70 ribosomal S6 kinase, p90RSK,serum and glucocorticoid-regulated kinase (SGK), and protein kinase C.The upstream kinase phosphorylating the regulatory site of Akt near thecarboxy terminus has not been identified yet, but recent reports imply arole for the integrin-inked kinase (ILK-1), a serine/threonine proteinkinase, or autophosphorylation.

Analysis of Akt levels in human tumors showed that Akt2 is overexpressedin a significant number of ovarian (J. Q. Cheng et al. Proc. Natl. Acad.Sci. U.S.A. 89:9267-9271(1992)) and pancreatic cancers (J. Q. Cheng etal. Proc. Natl. Acad. Sci. U.S.A. 93:3636-3641 (1996)). Similarly, Akt3was found to be overexpressed in breast and prostate cancer cell lines(Nakatani et al. J. Biol. Chem. 274:21528-21532 (1999).

The tumor suppressor PTEN, a protein and lipid phosphatase thatspecifically removes the 3′ phosphate of PtdIns(3,4,5)-P3, is a negativeregulator of the PI3K/Akt pathway (Li et al. Science 275:1943-1947(1997), Stambolic et al. Cell 95:29-39 (1998), Sun et al. Proc. Natl.Acad. Sci. U.S.A. 96:6199-6204 (1999)). Germline mutations of PTEN areresponsible for human cancer syndromes such as Cowden disease (Liaw etal. Nature Genetics 16:64-67 (1997)). PTEN is deleted in a largepercentage of human tumors and tumor cell lines without functional PTENshow elevated levels of activated Akt (Li et al. supra, Guldberg et al.Cancer Research 57:3660-3663 (1997), Risinger et al. Cancer Research57:4736-4738 (1997)).

These observations demonstrate that the PI3K/Akt pathway plays importantroles for regulating cell survival or apoptosis in tumorigenesis.

Inhibition of Akt activation and activity can be achieved by inhibitingPI3K with inhibitors such as LY294002 and wortmannin. However, PI3Kinhibition has the potential to indiscriminately affect not just allthree Akt isozymes but also other PH domain-containing signalingmolecules that are dependent on PdtIns(3,4,5)-P3, such as the Tec familyof tyrosine kinases. Furthermore, it has been disclosed that Akt can beactivated by growth signals that are independent of PI3K.

Alternatively, Akt activity can be inhibited by blocking the activity ofthe upstream kinase PDK1. No specific PDK1 inhibitors have beendisclosed. Again, inhibition of PDK1 would result in inhibition ofmultiple protein kinases whose activities depend on PDK1, such asatypical PKC isoforms, SGK, and S6 kinases (Williams et al. Curr. Biol.10:439-448 (2000).

It is an object of the instant invention to provide novel compounds thatare inhibitors of Akt.

It is also an object of the present invention to provide pharmaceuticalcompositions that comprise the novel compounds that are inhibitors ofAkt.

It is also an object of the present invention to provide a method fortreating cancer that comprises administering such inhibitors of Aktactivity.

SUMMARY OF THE INVENTION

The instant invention provides for compounds that inhibit Akt activity.In particular, the compounds disclosed selectively inhibit one or two ofthe Akt isoforms. The invention also provides for compositionscomprising such inhibitory compounds and methods of inhibiting Aktactivity by administering the compound to a patient in need of treatmentof cancer.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the instant invention are useful in the inhibition ofthe activity of the serine/threonine kinase Akt. In a first embodimentof this invention, the inhibitors of Akt activity are illustrated by theFormula A:

wherein:

a is 0 or 1; b is 0 or 1; m is 0, 1 or 2; n is 0, 1, 2, 3or 4; p is 0,1, 2, 3, 4 or 5; and t is 2, 3, 4, 5 or 6;

u, v, w and x are independently selected from: CH and N;

y and z are independently selected from: CH and N, provided that atleast one of y and z is N;

Ring K is selected from: (C₃-C₈)cycloalkyl, aryl, heteroaryl andheterocyclyl, with the proviso that ring K is not phenyl;

R¹ is independently selected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)₂NR⁷R⁸, S(O)₂—(C₁-C₁₀)alkyl and(C═O)_(a)O_(b)-heterocyclyl, said alkyl, aryl, alkenyl, alkynyl,cycloalkyl, and heterocyclyl is optionally substituted with one or moresubstituents selected from R⁶;

R² is independently selected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)₂NR⁷R⁸, S(O)₂—(C₁-C₁₀)alkyl and(C═O)_(a)O_(b)-heterocyclyl, said alkyl, aryl, alkenyl, alkynyl,cycloalkyl, and heterocyclyl is optionally substituted with one or moresubstituents selected from R⁶;

R³ and R⁴ are independently selected from: H, (C₁-C₆)alkyl and(C₁-C₆)perfluoroalkyl, or R³ and R⁴ are combined to form —(CH₂)_(t)—wherein one of the carbon atoms is optionally replaced by a moietyselected from O, S(O)_(m), —N(R^(b))C(O)—, and —N(COR^(a))—;

R⁵ is selected from: NR⁷R⁸;

R⁶ is: (C═O)_(a)O_(b)C₁-C₁₀ alkyl, (C═O)_(a)O_(b)aryl, C₂-C₁₀ alkenyl,C₂-C₁₀ alkynyl, (C═O)_(a)O_(b) heterocyclyl, CO₂H, halo, CN, OH,O_(b)C₁-C₆ perfluoroalkyl, O_(a)(C═O)_(b)NR⁷R⁸, oxo, CHO, (N═O)R⁷R⁸,S(O)₂NR⁷R⁸, S(O)₂—(C₁-C₁₀)alkyl or (C═O)_(a)O_(b)C₃-C₈ cycloalkyl, saidalkyl, aryl, alkenyl, alkynyl, heterocyclyl, and cycloalkyl optionallysubstituted with one or more substituents selected from R^(6a);

R^(6a) is selected from: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,O_(a)(C₁-C₃)perfluoroalkyl, (C₀-C₆)alkylene-S(O)_(m)R^(a), oxo, OH,halo, CN, (C₂-C₁₀)alkenyl, (C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl,(C₀-C₆)alkylene-aryl, (C₀-C₆)alkylene-heterocyclyl,(C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a), (C₀-C₆)alkylene-CO₂R^(a), C(O)H,and (C₀-C₆)alkylene-CO₂H, said alkyl, alkenyl, alkynyl, cycloalkyl,aryl, and heterocyclyl is optionally substituted with up to threesubstituents selected from R^(6b);

R^(6b) is selected from: (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,O_(a)(C₁-C₃)perfluoroalkyl, (C₀-C₆)alkylene-S(O)_(m)R^(a), oxo, OH,halo, CN, (C₂-C₁₀)alkenyl, (C₂-C₁₀)alkynyl, (C₃-C₆)cycloalkyl,(C₀-C₆)alkylene-aryl, (C₀-C₆)alkylene-heterocyclyl,(C₀-C₆)alkylene-N(R^(b))₂, C(O)R^(a), (C₀-C₆)alkylene-CO₂R^(a), C(O)H,and (C₀-C₆)alkylene-CO₂H, said alkyl, alkenyl, alkynyl, cycloalkyl,aryl, and heterocyclyl is optionally substituted with up to threesubstituents selected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂;

R⁷ and R⁸ are independently selected from: H,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, (C═O)_(a)O_(b)(C₃-C₈)cycloalkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)-heterocyclyl, (C₂-C₁₀)alkenyl,(C₂-C₁₀)alkynyl, SO₂R^(a), and (C═O)_(a)NR^(b) ₂, said alkyl,cycloalkyl, aryl, heterocylyl, alkenyl, and alkynyl is optionallysubstituted with one or more substituents selected from R^(6a), or R⁷and R⁸ can be taken together with the nitrogen to which they areattached to form a monocyclic or bicyclic heterocycle with 3-7 membersin each ring and optionally containing, in addition to the nitrogen, oneor two additional heteroatoms selected from N, O and S, said monocyclicor bicyclic heterocycle optionally substituted with one or moresubstituents selected from R^(6a);

R^(a) is (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, aryl, or heterocyclyl; and

R^(b) is H, (C₁-C₆)alkyl, aryl, heterocyclyl, (C₃-C₆)cycloalkyl,(C═O)_(a)O_(b)(C₁-C₆)alkyl, or S(O)₂R^(a);

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a second embodiment of this invention, the inhibitors of Akt activityare illustrated by the Formula B:

wherein:

q is 0, 1, 2, 3 or 4;

Q is independently selected from: heterocyclyl, said heterocyclyloptionally substituted with one to three substituents selected fromR^(6b);

and all other substituents and variables are as defined in the firstembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a third embodiment the inhibitors of the instant invention areillustrated by the Formula C:

wherein:

all other substituents and variables are as defined in the secondembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a fourth embodiment the inhibitors of the instant invention areillustrated by the Formula D:

wherein:

all other substituents and variables are as defined in the secondembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a fifth embodiment the inhibitors of the instant invention areillustrated by the Formula E:

wherein:

all other substituents and variables are as defined in the secondembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a sixth embodiment the inhibitors of the instant invention areillustrated by the Formula F:

wherein:

all other substituents and variables are as defined in the firstembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In a seventh embodiment the inhibitors of the instant invention areillustrated by the Formula G:

wherein:

R³ and R⁴ are independently: H and (C₁-C₆)alkyl, wherein said alkyl isoptionally substituted with up to three substituents selected from: OHand halo; and wherein R³ and R⁴ may be joined to form a(C₃-C₇)cycloalkyl; and

all other substituents and variables are as defined in the firstembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

In an eighth embodiment the inhibitors of the instant invention areillustrated by the Formula G:

wherein:

R³ and R⁴ are independently: H and (C₁-C₆)alkyl; and

all other substituents and variables are as defined in the firstembodiment;

or a pharmaceutically acceptable salt or a stereoisomer thereof.

Specific compounds of the instant invention include:

-   5-Methoxy-2(4-{[4-5-pyridin-2-yl-4H-1,2,4-triazol)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridine    (1-7);-   2-(4-{[4-(5-Pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one    (1-8);-   1-{4-[4-(3-pyrimidin-5-ylquinoxalin-2-yl)benzyl]cyclohexyl}-1,3-dihydro-2H-benimidazol-2-one    (1-9);-   3-[3-(4-{[4-2-oxo-2,3-dihydro-1H-benimidazol-1-yl)cyclohexyl]methyl}phenyl)quinoxalin-2-yl]thiophene-2-carbaldehyde    (1-10);-   1-(4-{4-[3-(1H-pyrazol-5-yl)quinoxalin-2-yl]benzyl}cyclohexyl)-1,3-dihydro-2H-benzimidazol-2-one    (1-11); and-   2-[4-(1-amino-1-methylethyl)phenyl]-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one    (2-7);-   or a pharmaceutically acceptable salt or stereoisomer thereof.

Examples of the compounds of the instant invention include TFA salts ofthe following compounds:

-   5-Methoxy-2(4-{[4-5-pyridin-2-yl-4H-1,2,4-triazol)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridine    (1-7);-   2-(4-{[4-(5-Pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one    (1-8);-   1-{4-[4-(3-pyrimidin-5-ylquinoxalin-2-yl)benzyl]cyclohexyl}-1,3-dihydro-2H-benimidazol-2-one    (1-9);-   3-[3-(4-{[4-2-oxo-2,3-dihydro-1H-benimidazol-1-yl)cyclohexyl]methyl}phenyl)quinoxalin-2-yl]thiophene-2-carbaldehyde    (1-10); and-   1-(4-{4-[3-(1H-pyrazol-5-yl)quinoxalin-2-yl]benzyl}cyclohexyl)-1,3-dihydro-2H-benzimidazol-2-one    (1-11);-   or a stereoisomer thereof.

The compounds of the present invention may have asymmetric centers,chiral axes, and chiral planes (as described in: E. L. Eliel and S. H.Wilen, Stereochemistry of Carbon Compounds, John Wiley & Sons, New York,1994, pages 1119-1190), and occur as racemates, racemic mixtures, and asindividual diastereomers, with all possible isomers and mixturesthereof, including optical isomers, all such stereoisomers beingincluded in the present invention.

In addition, the compounds disclosed herein may exist as tautomers andboth tautomeric forms are intended to be encompassed by the scope of theinvention, even though only one tautomeric structure is depicted. Forexample, any claim to compound A below is understood to includetautomeric structure B, and vice versa, as well as mixtures thereof. Thetwo tautomeric forms of the benzimidazolonyl moiety are also within thescope of the instant invention.

Tetrazoles exist as a mixture of 1H/2H tautomers. The tautomeric formsof the tetrazol moiety are also within the scope of the instantinvention.

When any variable (e.g. R¹, R², R^(6b), etc.) occurs more than one timein any constituent, its definition on each occurrence is independent atevery other occurrence. Also, combinations of substituents and variablesare permissible only if such combinations result in stable compounds.Lines drawn into the ring systems from substituents represent that theindicated bond may be attached to any of the substitutable ring atoms.If the ring system is bicyclic, it is intended that the bond be attachedto any of the suitable atoms on either ring of the bicyclic moiety.

It is understood that substituents and substitution patterns on thecompounds of the instant invention can be selected by one of ordinaryskill in the art to provide compounds that are chemically stable andthat can be readily synthesized by techniques known in the art, as wellas those methods set forth below, from readily available startingmaterials. If a substituent is itself substituted with more than onegroup, it is understood that these multiple groups may be on the samecarbon or on different carbons, so long as a stable structure results.The phrase “optionally substituted with one or more substituents” shouldbe taken to be equivalent to the phrase “optionally substituted with atleast one substituent” and in such cases the preferred embodiment willhave from zero to four substituents, and the more preferred embodimentwill have from zero to three substituents.

As used herein, “alkyl” is intended to include both branched andstraight-chain saturated aliphatic hydrocarbon groups having thespecified number of carbon atoms. For example, C₁-C₁₀, as in“(C₁-C₁₀)alkyl” is defined to include groups having 1, 2, 3, 4, 5, 6, 7,8, 9 or 10 carbons in a linear or branched arrange-ment. For example,“(C₁-C₁₀)alkyl” specifically includes methyl, ethyl, n-propyl, i-propyl,n-butyl, t-butyl, i-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl,and so on.

The term “cycloalkyl” means a monocyclic saturated aliphatic hydrocarbongroup having the specified number of carbon atoms. For example,“cycloalkyl” includes cyclopropyl, methyl-cyclopropyl,2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl, and so on.“Alkoxy” represents either a cyclic or non-cyclic alkyl group ofindicated number of carbon atoms attached through an oxygen bridge.“Alkoxy” therefore encompasses the definitions of alkyl and cycloalkylabove.

If no number of carbon atoms is specified, the term “alkenyl” refers toa non-aromatic hydrocarbon radical, straight, branched or cyclic,containing from 2 to 10 carbon atoms and at least one carbon to carbondouble bond. Preferably one carbon to carbon double bond is present, andup to four non-aromatic carbon-carbon double bonds may be present. Thus,“(C₂-C₁₀)alkenyl” means an alkenyl radical having from 2 to 10 carbonatoms. Alkenyl groups include ethenyl, propenyl, butenyl,2-methylbutenyl and cyclohexenyl. The straight, branched or cyclicportion of the alkenyl group may contain double bonds and may besubstituted if a substituted alkenyl group is indicated.

The term “alkynyl” refers to a hydrocarbon radical straight, branched orcyclic, containing from 2 to 10 carbon atoms and at least one carbon tocarbon triple bond. Up to three carbon-carbon triple bonds may bepresent. Thus, “(C₂-C₁₀)alkynyl” means an alkynyl radical having from 2to 10 carbon atoms. Alkynyl groups include ethynyl, propynyl, butynyl,3-methylbutynyl and so on. The straight, branched or cyclic portion ofthe alkynyl group may contain triple bonds and may be substituted if asubstituted alkynyl group is indicated.

In certain instances, substituents may be defined with a range ofcarbons that includes zero, such as (C₀-C₆)alkylene-aryl. If aryl istaken to be phenyl, this definition would include phenyl itself as wellas —CH₂Ph, —CH₂CH₂Ph, CH(CH₃)CH₂CH(CH₃)Ph, and so on.

As used herein, “aryl” is intended to mean any stable monocyclic orbicyclic carbon ring of up to 7 atoms in each ring, wherein at least onering is aromatic. Examples of such aryl elements include phenyl,naphthyl, tetrahydro-naphthyl, indanyl and biphenyl. In cases where thearyl substituent is bicyclic and one ring is non-aromatic, it isunderstood that attachment is via the aromatic ring.

The term heteroaryl, as used herein, represents a stable monocyclic orbicyclic ring of up to 7 atoms in each ring, wherein at least one ringis aromatic and contains from 1 to 4 heteroatoms selected from the groupconsisting of O, N and S. Heteroaryl groups within the scope of thisdefinition include but are not limited to: acridinyl, carbazolyl,cinnolinyl, quinoxalinyl, pyrrazolyl, indolyl, benzotriazolyl, furanyl,thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl,oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl,pyrimidinyl, pyrrolyl, tetrahydroquinoline. As with the definition ofheterocycle below, “heteroaryl” is also understood to include theN-oxide derivative of any nitrogen-containing heteroaryl. In cases wherethe heteroaryl substituent is bicyclic and one ring is non-aromatic orcontains no heteroatoms, it is understood that attachment is via thearomatic ring or via the heteroatom containing ring, respectively. Suchheteraoaryl moieties for substituent Q include but are not limited to:2-benzimidazolyl, 2-quinolinyl, 3-quinolinyl, 4-quinolinyl,1-isoquinolinyl, 3-isoquinolinyl and 4-isoquinolinyl.

The term “heterocycle” or “heterocyclyl” as used herein is intended tomean a 3- to 10-membered aromatic or nonaromatic heterocycle containingfrom 1 to 4 heteroatoms selected from the group consisting of O, N andS, and includes bicyclic groups. “Heterocyclyl” therefore includes theabove mentioned heteroaryls, as well as dihydro and tetrathydro analogsthereof. Further examples of “heterocyclyl” include, but are not limitedto the following: benzoimidazolyl, benzoimidazolonyl, benzofuranyl,benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl,benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, imidazolyl,indolinyl, indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl,isoquinolyl, isothiazolyl, isoxazolyl, naphthpyridinyl, oxadiazolyl,oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl,pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl,pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl,tetrahydropyranyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl,thiazolyl, thienyl, triazolyl, azetidinyl, 1,4-dioxanyl,hexahydroazepinyl, piperazinyl, piperidinyl, pyridin-2-onyl,pyrrolidinyl, morpholinyl, thiomorpholinyl, dihydrobenzoimidazolyl,dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl,dihydrofuranyl, dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl,dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl,dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl, andN-oxides thereof. Attachment of a heterocyclyl substituent can occur viaa carbon atom or via a heteroatom.

As appreciated by those of skill in the art, “halo” or “halogen” as usedherein is intended to include chloro (Ch), fluoro (F), bromo (Br) andiodo (I).

In an embodiment, the moiety illustrated by the formula:

includes the following structures, which are meant to be merelyillustrative and not limiting; moreover any suitable carbon and/ornitrogen contained within the bicyclic structure may be optionallysubstituted with from one two three substituents selected from R¹:

In another embodiment, the moiety illustrated by the formula:

is selected from:

wherein any suitable carbon and/or nitrogen in the bicyclic structuremay be optionally substituted with from one to three substituentsselected from R¹.

In another embodiment, the moiety illustrated by the formula:

is selected from:

wherein any suitable carbon and/or nitrogen in the bicyclic structuremay be optionally substituted with from one to three substituentsselected from R¹.

In an embodiment, ring K is selected from heterocyclyl.

In an embodiment, ring K is selected from:

In an embodiment, n is 0, 1, 2 or 3.

In a further embodiment, n is 0, 1 or 2.

In another embodiment, n is 1.

In an embodiment, p is 0, 1, 2 or 3.

In a further embodiment, p is 0, 1 or 2.

In another embodiment, p is 1.

In an embodiment, y and z are N.

In another embodiment, y is N and z is CH.

In another embodiment, y is CH and z is N.

In an embodiment, Q is selected from: benzoimidazolyl,benzoimidazolonyl, benzofuranyl, benzofurazanyl, benzopyrazolyl,benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl,cinnolinyl, furanyl, imidazolyl, indolinyl, indolyl, indolazinyl,indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl,isoxazolyl, naphthpyridinyl, oxadiazolyl, oxazolyl, oxazoline,isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl,pyridopyridinyl, pyridazinyl, pyridyl, pyrimidyl, pyrrolyl,quinazolinyl, quinolyl, quinoxalinyl, tetrahydropyranyl, tetrazolyl,tetrazolopyridyl, thiadiazolyl, thiazolyl, thienyl, triazolyl,azetidinyl, 1,4-dioxanyl, hexahydroazepinyl, piperazinyl, piperidinyl,pyridin-2-onyl, pyrrolidinyl, morpholinyl, thiomorpholinyl,dihydrobenzoimidazolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl,dihydrobenzoxazolyl, dihydrofuranyl, dihydroimidazolyl, dihydroindolyl,dihydroisooxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl,dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl, andN-oxides thereof, optionally substituted with one to three substituentsselected from R^(6b). Attachment of a heterocyclyl substituent can occurvia a carbon atom or via a heteroatom.

In a further embodiment, Q is selected from: 2-azepinone,benzimidazolyl, benzimidazolonyl, 2-diazapinone, imidazolyl,2-imidazolidinone, indolyl, isoquinolinyl, morpholinyl, piperidyl,piperazinyl, pyridyl, pyrrolidinyl, 2-piperidinone, 2-pyrimidinone,2-pyrollidinone, quinolinyl, tetrazolyl, tetrahydrofuryl,tetrahydroisoquinolinyl, and thienyl, optionally substituted with one tothree substituents selected from R^(6b). Attachment of a heterocyclylsubstituent can occur via a carbon atom or via a heteroatom.

In still a further embodiment, when Q is heterocyclyl, Q is selectedfrom:

which are optionally substituted with one to three substituents selectedfrom R^(6b).

In yet a further embodiment, when Q is heterocyclyl, Q is selected from

which is optionally substituted with one substituent selected fromR^(6b).

In an embodiment, R¹ is selected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)₂NR⁷R⁸, S(O)₂—(C₁-C₁₀)alkyl and(C═O)_(a)O_(b)-heterocyclyl, said alkyl, aryl, alkenyl, alkynyl,cycloalkyl, and heterocyclyl is optionally substituted with R^(b), OH,(C₁-C₆)alkoxy, halogen, CO₂H, CN, O(C═O)(C₁-C₆)alkyl, oxo, andN(R^(b))₂.

In another embodiment, R¹ is selected from: oxo,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN, (C₁-C₆)alkoxy,O(C═O)(C₁-C₆)alkyl and N(R^(b))₂.

In another embodiment, R¹ is selected from: oxo and O(C₁-C₆)alkyl.

In an embodiment, R² is selected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl,(C═O)_(a)O_(b)-aryl, (C═O)_(a)O_(b)(C₂-C₁₀)alkenyl,(C═O)_(a)O_(b)(C₂-C₁₀)alkynyl, CO₂H, halo, OH,O_(b)(C₁-C₆)perfluoroalkyl, (C═O)_(a)NR⁷R⁸, CN,(C═O)_(a)O_(b)(C₃-C₈)cycloalkyl, S(O)₂NR⁷R⁸, S(O)₂—(C₁-C₁₀)alkyl and(C═O)_(a)O_(b)-heterocyclyl, said alkyl, aryl, alkenyl, alkynyl,cycloalkyl, and heterocyclyl is optionally substituted with R^(b), OH,(C₁-C₆)alkoxy, halogen, CO₂H, CN, O(C═O)(C₁-C₆)alkyl, oxo, andN(R^(b))₂.

In another embodiment, R² is selected from: oxo,(C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN, (C₁-C₆)alkoxy,O(C═O)(C₁-C₆)alkyl, (C₂-C₁₀)alkenyl and N(R^(b))₂, said alkenyloptionally substituted with oxo.

In an embodiment, R³ and R⁴ are selected from H and (C₁-C₆)alkyl.

In another embodiment, R³ and R⁴ are H.

In an embodiment, when Q is substituted with R^(6b), said R^(6b) isheterocyclyl optionally substituted with up to three substituentsselected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂.

In another embodiment, when Q is substituted with R^(6b), said R^(6b) isselected from:

optionally substituted with 1 to 3 substituents selected from oxo, OH,N(R^(b))₂, halogen and O_(a)(C₁-C₆)alkyl.

In an embodiment R^(b) is independently selected from H and(C₁-C₆)alkyl.

In an embodiment of Formula A, ring K is heterocyclyl; R¹ is selectedfrom: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN,(C₁-C₆)alkoxy, O(C═O)(C₁-C₆)alkyl and N(R^(b))₂; R² is selected from:oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN, (C₁-C₆)alkoxy,O(C═O)(C₁-C₆)alkyl, (C₂-C₁₀)alkenyl and N(R^(b))₂, said alkenyloptionally substituted with oxo; R³ and R⁴ are H; Q is independentlyselected from: heterocyclyl, said heterocyclyl optionally substitutedwith one to three substituents selected from R^(6b); R^(6b) isheterocyclyl optionally substituted with up to three substituentsselected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂; R^(b) is independentlyselected from H and (C₁-C₆)alkyl; all other substituents and variablesare as defined in Formula A above.

In an embodiment of Formula D, ring K is heterocyclyl; R¹ is selectedfrom: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN,(C₁-C₆)alkoxy, O(C═O)(C₁-C₆)alkyl and N(R^(b))₂; R² is selected from:oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN, (C₁-C₆)alkoxy,O(C═O)(C₁-C₆)alkyl, (C₂-C₁₀)alkenyl and N(R^(b))₂, said alkenyloptionally substituted with oxo; R³ and R⁴ are H; Q is independentlyselected from: heterocyclyl, said heterocyclyl optionally substitutedwith one to three substituents selected from R^(6b); R^(6b) isheterocyclyl optionally substituted with up to three substituentsselected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂; R^(b) is independentlyselected from H and (C₁-C₆)alkyl; all other substituents and variablesare as defined in Formula D above.

In an embodiment of Formula E, ring K is heterocyclyl; R¹ is selectedfrom: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN,(C₁-C₆)alkoxy, O(C═O)(C₁-C₆)alkyl and N(R^(b))₂; R² is selected from:oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN, (C₁-C₆)alkoxy,O(C═O)(C₁-C₆)alkyl, (C₂-C₁₀)alkenyl and N(R^(b))₂, said alkenyloptionally substituted with oxo; R³ and R⁴ are H; Q is independentlyselected from: heterocyclyl, said heterocyclyl optionally substitutedwith one to three substituents selected from R^(6b); R^(6b) isheterocyclyl optionally substituted with up to three substituentsselected from R^(b), OH, (C₁-C₆)alkoxy, halogen, CO₂H, CN,O_(a)(C═O)_(b)(C₁-C₆)alkyl, oxo, and N(R^(b))₂; R^(b) is independentlyselected from H and (C₁-C₆)alkyl; all other substituents and variablesare as defined in Formula E above.

In an embodiment of Formula G, ring K is selected from heterocyclyl.

In an embodiment of Formula G, p is 0 and ring K is selected from:

In an embodiment of Formula G, p is 0 and ring K is

Included in the instant invention is the free form of compounds ofFormula A, as well as the pharmaceutically acceptable salts andstereoisomers thereof. Some of the isolated specific compoundsexemplified herein are the protonated salts of amine compounds. The term“free form” refers to the amine compounds in non-salt form. Theencompassed pharmaceutically acceptable salts not only include theisolated salts exemplified for the specific compounds described herein,but also all the typical pharmaceutically acceptable salts of the freeform of compounds of Formula A. The free form of the specific saltcompounds described may be isolated using techniques known in the art.For example, the free form may be regenerated by treating the salt witha suitable dilute aqueous base solution such as dilute aqueous NaOH,potassium carbonate, ammonia and sodium bicarbonate. The free forms maydiffer from their respective salt forms somewhat in certain physicalproperties, such as solubility in polar solvents, but the acid and basesalts are otherwise pharmaceutically equivalent to their respective freeforms for purposes of the invention.

The pharmaceutically acceptable salts of the instant compounds can besynthesized from the compounds of this invention which contain a basicor acidic moiety by conventional chemical methods. Generally, the saltsof the basic compounds are prepared either by ion exchangechromatography or by reacting the free base with stoichiometric amountsor with an excess of the desired salt-forming inorganic or organic acidin a suitable solvent or various combinations of solvents. Similarly,the salts of the acidic compounds are formed by reactions with theappropriate inorganic or organic base.

Thus, pharmaceutically acceptable salts of the compounds of thisinvention include the conventional non-toxic salts of the compounds ofthis invention as formed by reacting a basic instant compound with aninorganic or organic acid. For example, conventional non-toxic saltsinclude those derived from inorganic acids such as hydrochloric,hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like, aswell as salts prepared from organic acids such as acetic, propionic,succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic,pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic,salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic,methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic(TFA) and the like.

When the compound of the present invention is acidic, suitable“pharmaceutically acceptable salts” refers to salts prepared formpharmaceutically acceptable non-toxic bases including inorganic basesand organic bases. Salts derived from inorganic bases include aluminum,ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganicsalts, manganous, potassium, sodium, zinc and the like. Particularlypreferred are the ammonium, calcium, magnesium, potassium and sodiumsalts. Salts derived from pharmaceutically acceptable organic non-toxicbases include salts of primary, secondary and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, such as arginine, betainecaffeine, choline, N,N¹-dibenzylethylenediamine, diethylamin,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylaminetripropylamine, tromethamine and the like.

The preparation of the pharmaceutically acceptable salts described aboveand other typical pharmaceutically acceptable salts is more fullydescribed by Berg et al., “Pharmaceutical Salts,” J. Pharm. Sci.,1977:66:1-19.

It will also be noted that the compounds of the present invention arepotentially internal salts or zwitterions, since under physiologicalconditions a deprotonated acidic moiety in the compound, such as acarboxyl group, may be anionic, and this electronic charge might then bebalanced off internally against the cationic charge of a protonated oralkylated basic moiety, such as a quaternary nitrogen atom.

UTILITY

The compounds of the instant invention are inhibitors of the activity ofAkt and are thus useful in the treatment of cancer, in particularcancers associated with irregularities in the activity of Akt anddownstream cellular targets of Akt. Such cancers include, but are notlimited to, ovarian, pancreatic, breast and prostate cancer, as well ascancers (including glioblastoma) where the tumor suppressor PTEN ismutated (Cheng et al., Proc. Natl. Acad. Sci. (1992) 89:9267-9271; Chenget al., Proc. Natl. Acad. Sci. (1996) 93:3636-3641; Bellacosa et al.,Int. J. Cancer (1995) 64:280-285; Nakatani et al., J. Biol. Chem. (1999)274:21528-21532; Graff, Expert. Opin. Ther. Targets (2002) 6(1):103-113;and Yamada and Araki, J. Cell Science. (2001) 114:2375-2382; Mischel andCloughesy, Brain Pathol. (2003) 13(1):52-61).

The compounds, compositions and methods provided herein are particularlydeemed useful for the treatment of cancer. Cancers that may be treatedby the compounds, compositions and methods of the invention include, butare not limited to: Cardiac: sarcoma (angiosarcoma, fibrosarcoma,rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma andteratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiatedsmall cell, undifferentiated large cell, adenocarcinoma), alveolar(bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma,chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus(squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma),stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductaladenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors,vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors,Karposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma,fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma,hamartoma, leiomyoma); Genitourinary tract: kidney (adenocarcinoma,Wilm's tumor [nephroblastoma], lymphoma, leukemia), bladder and urethra(squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma),prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma,embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma,interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors,lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma,hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bone:osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibroushistiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma(reticulum cell sarcoma), multiple myeloma, malignant giant cell tumorchordoma, osteochronfroma (osteocartilaginous exostoses), benignchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma andgiant cell tumors; Nervous system: skull (osteoma, hemangioma,granuloma, xanthoma, osteitis deformans), meninges (meningioma,meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma,glioma, ependymoma, germinoma [pinealoma], glioblastoma multiform,oligodendroglioma, schwannoma, retinoblastoma, congenital tumors),spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological:uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumorcervical dysplasia), ovaries (ovarian carcinoma [serouscystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiedcarcinoma], granulosa-thecal cell tumors, Sertoli-Leydig cell tumors,dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma,intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma),vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma(embryonal rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic:blood (myeloid leukemia [acute and chronic], acute lymphoblasticleukemia, chronic lymphocytic leukemia, myeloproliferative diseases,multiple myeloma, myelodysplastic syndrome), Hodgkin's disease,non-Hodgkin's lymphoma [malignant lymphoma]; Skin: malignant melanoma,basal cell carcinoma, squamous cell carcinoma, Karposi's sarcoma, molesdysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis;and Adrenal glands: neuroblastoma. Thus, the term “cancerous cell” asprovided herein, includes a cell afflicted by any one of theabove-identified conditions.

Cancers that may be treated by the compounds, compositions and methodsof the invention include, but are not limited to: breast, prostate,colon, lung, brain, testicular, stomach, pancrease, skin, smallintestine, large intestine, throat, head and neck, oral, bone, liver,bladder, kidney, thyroid and blood.

Akt signaling regulates multiple critical steps in angiogenesis.Shiojima and Walsh, Circ. Res. (2002) 90:1243-1250. The utility ofangiogenesis inhibitors in the treatment of cancer is known in theliterature, see J. Rak et al. Cancer Research, 55:4575-4580, 1995 andDredge et al., Expert Opin. Biol. Ther. (2002) 2(8):953-966, forexample. The role of angiogenesis in cancer has been shown in numeroustypes of cancer and tissues: breast carcinoma (G. Gasparini and A. L.Harris, J. Clin. Oncol., 1995, 13:765-782; M. Toi et al., Japan. J.Cancer Res., 1994, 85:1045-1049); bladder carcinomas (A. J. Dickinson etal., Br. J. Urol., 1994, 74:762-766); colon carcinomas (L. M. Ellis etal., Surgery, 1996, 120(5):871-878); and oral cavity tumors (J. K.Williams et al., Am. J. Surg., 1994, 168:373-380). Other cancersinclude, advanced tumors, hairy cell leukemia, melanoma, advanced headand neck, metastatic renal cell, non-Hodgkin's lymphoma, metastaticbreast, breast adenocarcinoma, advanced melanoma, pancreatic, gastric,glioblastoma, lung, ovarian, non-small cell lung, prostate, small celllung, renal cell carcinoma, various solid tumors, multiple myeloma,metastatic prostate, malignant glioma, renal cancer, lymphoma,refractory metastatic disease, refractory multiple myeloma, cervicalcancer, Kaposi's sarcoma, recurrent anaplastic glioma, and metastaticcolon cancer (Dredge et al., Expert Opin. Biol. Ther. (2002)2(8):953-966). Thus, the Akt inhibitors disclosed in the instantapplication are also useful in the treatment of these angiogenesisrelated cancers.

Tumors which have undergone neovascularization show an increasedpotential for metastasis. In fact, angiogenesis is essential for tumorgrowth and metastasis. (S. P. Cunningham, et al., Can. Research, 61:3206-3211 (2001)). The Akt inhibitors disclosed in the presentapplication are therefore also useful to prevent or decrease tumor cellmetastasis.

Further included within the scope of the invention is a method oftreating or preventing a disease in which angiogenesis is implicated,which is comprised of administering to a mammal in need of suchtreatment a therapeutically effective amount of a compound of thepresent invention. Ocular neovascular diseases are an example ofconditions where much of the resulting tissue damage can be attributedto aberrant infiltration of blood vessels in the eye (see WO 00/30651,published 2 Jun. 2000). The undesirable infiltration can be triggered byischemic retinopathy, such as that resulting from diabetic retinopathy,retinopathy of prematurity, retinal vein occlusions, etc., or bydegenerative diseases, such as the choroidal neovascularization observedin age-related macular degeneration. Inhibiting the growth of bloodvessels by administration of the present compounds would thereforeprevent the infiltration of blood vessels and prevent or treat diseaseswhere angiogenesis is implicated, such as ocular diseases like retinalvascularization, diabetic retinopathy, age-related macular degeneration,and the like.

Further included within the scope of the invention is a method oftreating or preventing a non-malignant disease in which angiogenesis isimplicated, including but not limited to: ocular diseases (such as,retinal vascularization, diabetic retinopathy and age-related maculardegeneration), atherosclerosis, arthritis, psoriasis, obesity andAlzheimer's disease (Dredge et al., Expert Opin. Biol. Ther. (2002)2(8):953-966). In another embodiment, a method of treating or preventinga disease in which angiogenesis is implicated includes: ocular diseases(such as, retinal vascularization, diabetic retinopathy and age-relatedmacular degeneration), atherosclerosis, arthritis and psoriasis.

Further included within the scope of the invention is a method oftreating hyperproliferative disorders such as restenosis, inflammation,autoimmune diseases and allergy/asthma.

Further included within the scope of the instant invention is the use ofthe instant compounds to coat stents and therefore the use of theinstant compounds on coated stents for the treatment and/or preventionof restenosis (WO03/032809).

Further included within the scope of the instant invention is the use ofthe instant compounds for the treatment and/or prevention ofosteoarthritis (WO03/035048).

Further included within the scope of the invention is a method oftreating hyperinsulinism.

The compounds of the invention are also useful in preparing a medicamentthat is useful in treating the diseases described above, in particularcancer.

In an embodiment of the invention, the instant compound is a selectiveinhibitor whose inhibitory efficacy is dependent on the PH domain. Inthis embodiment, the compound exhibits a decrease in iii vitroinhibitory activity or no in vitro inhibitory activity against truncatedAkt proteins lacking the PH domain.

In a further embodiment, the instant compound is selected from the groupof a selective inhibitor of Akt1, a selective inhibitor of Akt2 and aselective inhibitor of both Akt1 and Akt2.

In another embodiment, the instant compound is selected from the groupof a selective inhibitor of Akt1, a selective inhibitor of Akt2, aselective inhibitor of Akt3 and a selective inhibitor of two of thethree Akt isoforms.

In another embodiment, the instant compound is a selective inhibitor ofall three Akt isoforms, but is not an inhibitor of one, two or all ofsuch Akt isoforms that have been modified to delete the PH domain, thehinge region or both the PH domain and the hinge region.

The present invention is further directed to a method of inhibiting Aktactivity which comprises administering to a mammal in need thereof apharmaceutically effective amount of the instant compound.

The compounds of this invention may be administered to mammals,including humans, either alone or, in combination with pharmaceuticallyacceptable carriers, excipients or diluents, in a pharmaceuticalcomposition, according to standard pharmaceutical practice. Thecompounds can be administered orally or parenterally, including theintravenous, intramuscular, intraperitoneal, subcutaneous, rectal andtopical routes of administration.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions and suchcompositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as calcium carbonate, sodium carbonate,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, microcrystalline cellulose, sodiumcrosscarmellose, corn starch, or alginic acid; binding agents, forexample starch, gelatin, polyvinyl-pyrrolidone or acacia, andlubricating agents, for example, magnesium stearate, stearic acid ortalc. The tablets may be uncoated or they may be coated by knowntechniques to mask the unpleasant taste of the drug or delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a watersoluble taste masking material such as hydroxypropylmethyl-cellulose orhydroxypropylcellulose, or a time delay material such as ethylcellulose, cellulose acetate buryrate may be employed.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with watersoluble carrier such as polyethyleneglycol or an oil medium, for examplepeanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active material in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethylene-oxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose, saccharin or aspartame.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as butylated hydroxyanisol or alpha-tocopherol.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present. These compositions may be preserved by theaddition of an anti-oxidant such as ascorbic acid.

The pharmaceutical compositions of the invention may also be in the formof an oil-in-water emulsion. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring phosphatides, for example soy bean lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening, flavouring agents, preservatives and antioxidants.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative, flavoring and coloring agentsand antioxidant.

The pharmaceutical compositions may be in the form of sterile injectableaqueous solutions. Among the acceptable vehicles and solvents that maybe employed are water, Ringer's solution and isotonic sodium chloridesolution.

The sterile injectable preparation may also be a sterile injectableoil-in-water microemulsion where the active ingredient is dissolved inthe oily phase. For example, the active ingredient may be firstdissolved in a mixture of soybean oil and lecithin. The oil solutionthen introduced into a water and glycerol mixture and processed to forma microemulation.

The injectable solutions or microemulsions may be introduced into apatient's blood-stream by local bolus injection. Alternatively, it maybe advantageous to administer the solution or microemulsion in such away as to maintain a constant circulating concentration of the instantcompound. In order to maintain such a constant concentration, acontinuous intravenous delivery device may be utilized. An example ofsuch a device is the Deltec CADD-PLUS™ model 5400 intravenous pump.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension for intramuscular andsubcutaneous administration. This suspension may be formulated accordingto the known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example as a solution in 1,3-butane diol. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose any bland fixed oil may be employed includingsynthetic mono- or diglycerides. In addition, fatty acids such as oleicacid find use in the preparation of injectables.

Compounds of Formula A may also be administered in the form ofsuppositories for rectal administration of the drug. These compositionscan be prepared by mixing the drug with a suitable non-irritatingexcipient which is solid at ordinary temperatures but liquid at therectal temperature and will therefore melt in the rectum to release thedrug. Such materials include cocoa butter, glycerinated gelatin,hydrogenated vegetable oils, mixtures of polyethylene glycols of variousmolecular weights and fatty acid esters of polyethylene glycol.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compound of Formula A are employed. (For purposesof this application, topical application shall include mouth washes andgargles.)

The compounds for the present invention can be administered inintranasal form via topical use of suitable intranasal vehicles anddelivery devices, or via transdermal routes, using those forms oftransdermal skin patches well known to those of ordinary skill in theart. To be administered in the form of a transdermal delivery system,the dosage administration will, of course, be continuous rather thanintermittent throughout the dosage regimen. Compounds of the presentinvention may also be delivered as a suppository employing bases such ascocoa butter, glycerinated gelatin, hydrogenated vegetable oils,mixtures of polyethylene glycols of various molecular weights and fattyacid esters of polyethylene glycol.

When a composition according to this invention is administered into ahuman subject, the daily dosage will normally be determined by theprescribing physician with the dosage generally varying according to theage, weight, and response of the individual patient, as well as theseverity of the patient's symptoms.

In an embodiment, a suitable amount of an inhibitor of Akt isadministered to a mammal undergoing treatment for cancer. Administrationoccurs in an amount of inhibitor of between about 0.1 mg/kg of bodyweight to about 60 mg/kg of body weight per day, or between 0.5 mg/kg ofbody weight to about 40 mg/kg of body weight per day. Anothertherapeutic dosage that comprises the instant composition includes fromabout 0.01 mg to about 1000 mg of inhibitor of Akt. In anotherembodiment, the dosage comprises from about 1 mg to about 1000 mg ofinhibitor of Akt.

The instant compounds are also useful in combination with knowntherapeutic agents and anti-cancer agents. For example, instantcompounds are useful in combination with known anti-cancer agents.Combinations of the presently disclosed compounds with other anti-canceror chemotherapeutic agents are within the scope of the invention.Examples of such agents can be found in Cancer Principles and Practiceof Oncology by V. T. Devita and S. Hellman (editors), 6^(th) edition(Feb. 15, 2001), Lippincott Williams & Wilkins Publishers. A person ofordinary skill in the art would be able to discern which combinations ofagents would be useful based on the particular characteristics of thedrugs and the cancer involved. Such anti-cancer agents include thefollowing: estrogen receptor modulators, androgen receptor modulators,retinoid receptor modulators, cytotoxic/cytostatic agents,antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoAreductase inhibitors and other angiogenesis inhibitors, HIV proteaseinhibitors, reverse transcriptase inhibitors, inhibitors of cellproliferation and survival signaling, and agents that interfere withcell cycle checkpoints. The instant compounds are particularly usefulwhen co-administered with radiation therapy.

“Estrogen receptor modulators” refers to compounds that interfere withor inhibit the binding of estrogen to the receptor, regardless ofmechanism. Examples of estrogen receptor modulators include, but are notlimited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081,toremifene, fulvestrant,4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]phenyl]-2H-1-benzopyran-3-yl]-phenyl-2,2-dimethylpropanoate,4,4′-dihydroxybenzophenone-2,4-dinitrophenyl-hydrazone, and SH646.

“Androgen receptor modulators” refers to compounds which interfere orinhibit the binding of androgens to the receptor, regardless ofmechanism. Examples of androgen receptor modulators include finasterideand other 5α-reductase inhibitors, nilutamide, flutamide, bicalutamide,liarozole, and abiraterone acetate.

“Retinoid receptor modulators” refers to compounds which interfere orinhibit the binding of retinoids to the receptor, regardless ofmechanism. Examples of such retinoid receptor modulators includebexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid,α-difluoromethylornithine, ILX23-7553, trans-N-(4′-hydroxyphenyl)retinamide, and N-4-carboxyphenyl retinamide.

“Cytotoxic/cytostatic agents” refer to compounds which cause cell deathor inhibit cell proliferation primarily by interfering directly with thecell's functioning or inhibit or interfere with cell myosis, includingalkylating agents, tumor necrosis factors, intercalators, hypoxiaactivatable compounds, microtubule inhibitors/microtubule-stabilizingagents, inhibitors of mitotic kinesins, histone deacetylase inhibitors,inhibitors of kinases involved in mitotic progression, inhibitors ofkinases involved in growth factor and cytokine signal transductionpathways, antimetabolites, biological response modifiers,hormonal/anti-hormonal therapeutic agents, haematopoietic growthfactors, monoclonal antibody targeted therapeutic agents, topoisomeraseinhibitors, proteosome inhibitors, ubiquitin ligase inhibitors, andaurora kinase inhibitors.

Examples of cytotoxic/cytostatic agents include, but are not limited to,sertenef, cachectin, ifosfamide, tasonermin, lonidamine, carboplatin,altretamine, prednimustine, dibromodulcitol, ranimustine, fotemustine,nedaplatin, oxaliplatin, temozolomide, heptaplatin, estramustine,improsulfan tosilate, trofosfamide, nimustine, dibrospidium chloride,pumitepa, lobaplatin, satraplatin, profiromycin, cisplatin, irofulven,dexifosfamide, cis-aminedichloro(2-methyl-pyridine)platinum,benzylguanine, glufosfamide, GPX100, (trans, trans,trans)-bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]bis[diamine(chloro)platinum(II)]tetrachloride, diarizidinylspermine, arsenic trioxide,1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin,idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin,pinafide, valrubicin, amrubicin, antineoplaston,3′-deamino-3′-morpholino-13-deoxo-10-hydroxycarminomycin, annamycin,galarubicin, elinafide, MEN10755,4-demethoxy-3-deamino-3-aziridinyl-4-methylsulphonyl-daunorubicin (seeWO 00/50032), Raf kinase inhibitors (such as Bay43-9006) and mTORinhibitors (such as Wyeth's CCI-779).

An example of a hypoxia activatable compound is tirapazamine.

Examples of proteosome inhibitors include but are not limited tolactacystin and MLN-341 (Velcade).

Examples of microtubule inhibitors/microtubule-stabilising agentsinclude paclitaxel, vindesine sulfate,3′,4′-didehydro-4′-deoxy-8′-norvincaleukoblastine, docetaxol, rhizoxin,dolastatin, mivobulin isethionate, auristatin, cemadotin, RPR109881,BMS184476, vinflunine, cryptophycin,2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl) benzene sulfonamide,anhydrovinblastine,N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butylamide,TDX258, the epothilones (see for example U.S. Pat. Nos. 6,284,781 and6,288,237) and BMS188797. In an embodiment the epothilones are notincluded in the microtubule inhibitors/microtubule-stabilising agents.

Some examples of topoisomerase inhibitors are topotecan, hycaptamine,irinotecan, rubitecan,6-ethoxypropionyl-3′,4′-O-exo-benzylidene-chartreusin,9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-k1]acridine-2-(6H)propanamine, 1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3′,4′:b,7]-indolizino[1,2b]quinoline-10,13(9H,15H)dione,lurtotecan, 7-[2-(N-isopropylamino)ethyl]-(20S)camptothecin, BNP1350,BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane,2′-dimethylamino-2′-deoxy-etoposide, GL331,N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazole-1-carboxamide,asulacrine, (5a, 5aB,8aa,9b)-9-[2-[N-[2-(dimethylamino)ethyl]-N-methylamino]ethyl]-5-[4-hydroOxy-3,5-dimethoxyphenyl]-5,5a,6,8,8a,9-hexohydrofuro(3′,4′:6,7)naphtho(2,3-d)-1,3-dioxol-6-one,2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]-phenanthridinium,6,9-bis [(2-aminoethyl)amino]benzo[g]isoguinoline-5,10-dione,5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-pyrazolo[4,5,1-de]acridin-6-one,N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl]formamide,N-(2-(dimethylamino)ethyl)acridine-4-carboxamide,6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno[2,1-c]quinolin-7-one,and dimesna.

Examples of inhibitors of mitotic kinesins, and in particular the humanmitotic kinesin KSP, are described in PCT Publications WO 01/30768, WO01/98278, WO 03/049527, WO 03/049679, WO 03/050064, WO 03/050122, WO03/049678 and WO 03/039460. In an embodiment inhibitors of mitotickinesins include, but are not limited to inhibitors of KSP, inhibitorsof MKLP1, inhibitors of CENP-E, inhibitors of MCAK and inhibitors ofRab6-KIFL.

Examples of “histone deacetylase inhibitors” include, but are notlimited to, SAHA, TSA, oxamflatin, PXD101, MG98 and scriptaid. Furtherreference to other histone deacetylase inhibitors may be found in thefollowing manuscript; Miller, T. A. et al. J. Med. Chem.46(24):5097-5116 (2003).

“Inhibitors of kinases involved in mitotic progression” include, but arenot limited to, inhibitors of aurora kinase, inhibitors of Polo-likekinases (PLK; in particular inhibitors of PLK-1), inhibitors of bub-1and inhibitors of bub-R1. An example of an “aurora kinase inhibitor” isVX-680.

“Antiproliferative agents” includes antisense RNA and DNAoligonucleotides such as G3139, ODN698, RVASKRAS, GEM231, and INX3001,and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin,doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine,cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed,paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed, pemetrexed,nelzarabine, 2′-deoxy-2′-methylidenecytidine,2′-fluoromethylene-2′-deoxycytidine,N-[5-(2,3-dihydro-benzofuryl)sulfonyl]-N′-(3,4-dichlorophenyl)urea,N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero-B-L-manno-heptopyranosyl]adenine,aplidine, ecteinascidin, troxacitabine,4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][1,4]thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamicacid, aminopterin, 5-flurouracil, alanosine,11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-14-oxa-1,11-diazatetracyclo(7.4.1.0.0)-tetradeca-2,4,6-trien-9-ylacetic acid ester, swainsonine, lometrexol, dexrazoxane, methioninase,2′-cyano-2′-deoxy-N4-palmitoyl-1-B-D-arabino furanosyl cytosine,3-aminopyridine-2-carboxaldehyde thiosemicarbazone and trastuzumab.

Examples of monoclonal antibody targeted therapeutic agents includethose therapeutic agents which have cytotoxic agents or radioisotopesattached to a cancer cell specific or target cell specific monoclonalantibody. Examples include Bexxar.

“HMG-CoA reductase inhibitors” refers to inhibitors of3-hydroxy-3-methylglutaryl-CoA reductase. Examples of HMG-CoA reductaseinhibitors that may be used include but are not limited to lovastatin(MEVACOR®; see U.S. Pat. Nos. 4,231,938, 4,294,926 and 4,319,039),simvastatin (ZOCOR®; see U.S. Pat. Nos. 4,444,784, 4,820,850 and4,916,239), pravastatin (PRAVACHOL®; see U.S. Pat. Nos. 4,346,227,4,537,859, 4,410,629, 5,030,447 and 5,180,589), fluvastatin (LESCOL®;see U.S. Pat. Nos. 5,354,772, 4,911,165, 4,929,437, 5,189,164,5,118,853, 5,290,946 and 5,356,896), atorvastatin (LIPITOR®; see U.S.Pat. Nos. 5,273,995, 4,681,893, 5,489,691 and 5,342,952) andcerivastatin (also known as rivastatin and BAYCHOL®; see U.S. Pat. No.5,177,080). The structural formulas of these and additional HMG-CoAreductase inhibitors that may be used in the instant methods aredescribed at page 87 of M. Yalpani, “Cholesterol Lowering Drugs”,Chemistry & Industry, pp. 85-89 (5 Feb. 1996) and U.S. Pat. Nos.4,782,084 and 4,885,314. The term HMG-CoA reductase inhibitor as usedherein includes all pharmaceutically acceptable lactone and open-acidforms (i.e., where the lactone ring is opened to form the free acid) aswell as salt and ester forms of compounds which have HMG-CoA reductaseinhibitory activity, and therefor the use of such salts, esters,open-acid and lactone forms is included within the scope of thisinvention.

“Prenyl-protein transferase inhibitor” refers to a compound whichinhibits any one or any combination of the prenyl-protein transferaseenzymes, including farnesyl-protein transferase (FPTase),geranylgeranyl-protein transferase type I (GGPTase-I), andgeranylgeranyl-protein transferase type-II (GGPTase-II, also called RabGGPTase).

Examples of prenyl-protein transferase inhibitors can be found in thefollowing publications and patents: WO 96/30343, WO 97/18813, WO97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98/29119, WO95/32987, U.S. Pat. Nos. 5,420,245, 5,523,430, 5,532,359, 5,510,510,5,589,485, 5,602,098, European Patent Publ. 0 618 221, European PatentPubl. 0 675 112, European Patent Publ. 0 604 181, European Patent Publ.0 696 593, WO 94/19357, WO 95/08542, WO 95/11917, WO 95/12612, WO95/12572, WO 95/10514, U.S. Pat. No. 5,661,152, WO 95/10515, WO95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO96/22278, WO 96/24611, WO 96/24612, WO 96/05168, WO 96/05169, WO96/00736, U.S. Pat. No. 5,571,792, WO 96/17861, WO 96/33159, WO96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO96/30363, WO 96/31111, WO 96/31477, WO 96/31478, WO 96/31501, WO97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO98/02436, and U.S. Pat. No. 5,532,359. For an example of the role of aprenyl-protein transferase inhibitor on angiogenesis see European J. ofCancer, Vol. 35, No. 9, pp. 1394-1401 (1999).

“Angiogenesis inhibitors” refers to compounds that inhibit the formationof new blood vessels, regardless of mechanism. Examples of angiogenesisinhibitors include, but are not limited to, tyrosine kinase inhibitors,such as inhibitors of the tyrosine kinase receptors Flt-1 (VEGFR1) andFlk-1/KDR (VEGFR2), inhibitors of epidermal-derived, fibroblast-derived,or platelet derived growth factors, MMP (matrix metalloprotease)inhibitors, integrin blockers, interferon-α, interleukin-12, pentosanpolysulfate, cyclooxygenase inhibitors, including nonsteroidalanti-inflammatories (NSAIDs) like aspirin and ibuprofen as well asselective cyclooxy-genase-2 inhibitors like celecoxib and rofecoxib(PNAS, Vol. 89, p. 7384 (1992); JNCI, Vol. 69, p. 475 (1982); Arch.Opthalmol., Vol. 108, p. 573 (1990); Anat. Rec., Vol. 238, p. 68 (1994);FEBS Letters, Vol. 372, p. 83 (1995); Clin, Orthop. Vol. 313, p. 76(1995); J. Mol. Endocrinol., Vol. 16, p. 107 (1996); Jpn. J. Pharmacol.,Vol. 75, p. 105 (1997); Cancer Res., Vol. 57, p. 1625 (1997); Cell, Vol.93, p. 705 (1998); Intl. J. Mol. Med., Vol. 2, p. 715 (1998); J. Biol.Chem., Vol. 274, p. 9116 (1999)), steroidal anti-inflammatories (such ascorticosteroids, mineralocorticoids, dexamethasone, prednisone,prednisolone, methylpred, betamethasone), carboxyamidotriazole,combretastatin A-4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol,thalidomide, angiostatin, troponin-1, angiotensin II antagonists (seeFernandez et al., J. Lab. Clin. Med. 105:141-145 (1985)), and antibodiesto VEGF (see, Nature Biotechnology, Vol. 17, pp. 963-968 (October 1999);Kim et al., Nature, 362, 841-844 (1993); WO 00/44777; and WO 00/61186).

Other therapeutic agents that modulate or inhibit angiogenesis and mayalso be used in combination with the compounds of the instant inventioninclude agents that modulate or inhibit the coagulation and fibrinolysissystems (see review in Clin. Chem. La. Med. 38:679-692 (2000)). Examplesof such agents that modulate or inhibit the coagulation and fibrinolysispathways include, but are not limited to, heparin (see Thromb. Haemost.80:10-23 (1998)), low molecular weight heparins and carboxypeptidase Uinhibitors (also known as inhibitors of active thrombin activatablefibrinolysis inhibitor [TAFIa]) (see Thrombosis Res. 101:329-354(2001)). TAFIa inhibitors have been described in WO 03/13526.

“Agents that interfere with cell cycle checkpoints” refer to compoundsthat inhibit protein kinases that transduce cell cycle checkpointsignals, thereby sensitizing the cancer cell to DNA damaging agents.Such agents include inhibitors of ATR, ATM, the Chk1 and Chk2 kinasesand cdk and cdc kinase inhibitors and are specifically exemplified by7-hydroxystaurosporin, flavopiridol, CYC202 (Cyclacel) and BMS-387032.

“Inhibitors of cell proliferation and survival signaling pathway” referto compounds that inhibit signal transduction cascades downstream ofcell surface receptors. Such agents include inhibitors ofserine/threonine kinases (including but not limited to inhibitors of Aktsuch as described in WO 02/083064, WO 02/083139, WO 02/083140, WO02/083138, WO 03/086279, WO 03/086394, WO 03/086403, WO 03/086404 and WO04/041162), inhibitors of Raf kinase (for example BAY-43-9006),inhibitors of MEK (for example CI-1040 and PD-098059), inhibitors ofmTOR (for example Wyeth CCI-779), and inhibitors of PI3K (for exampleLY294002).

As described above, the combinations with NSAID's are directed to theuse of NSAID's which are potent COX-2 inhibiting agents. For purposes ofthis specification an NSAID is potent if it possesses an IC₅₀ for theinhibition of COX-2 of 1 μM or less as measured by cell or microsomalassays.

The invention also encompasses combinations with NSAID's which areselective COX-2 inhibitors. For purposes of this specification NSAID'swhich are selective inhibitors of COX-2 are defined as those whichpossess a specificity for inhibiting COX-2 over COX-1 of at least 100fold as measured by the ratio of IC₅₀ for COX-2 over IC₅₀ for COX-1evaluated by cell or microsomal assays. Such compounds include, but arenot limited to those disclosed in U.S. Pat. Nos. 5,474,995, 5,861,419,6,001,843, 6,020,343, 5,409,944, 5,436,265, 5,536,752, 5,550,142,5,604,260, 5,698,584, 5,710,140, WO 94/15932, U.S. Pat. Nos. 5,344,991,5,134,142, 5,380,738, 5,393,790, 5,466,823, 5,633,272 and 5,932,598, allof which are hereby incorporated by reference.

Inhibitors of COX-2 that are particularly useful in the instant methodof treatment are: 3-phenyl-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone;and5-chloro-3-(4-methylsulfonyl)phenyl-2-(2-methyl-5-pyridinyl)pyridine; ora pharmaceutically acceptable salt thereof.

Compounds that have been described as specific inhibitors of COX-2 andare therefore useful in the present invention include, but are notlimited to, the following: parecoxib, BEXTRA® and CELEBREX® or apharmaceutically acceptable salt thereof.

Other examples of angiogenesis inhibitors include, but are not limitedto, endostatin, ukrain, ranpirnase, IM862,5-methoxy-4-[2-methyl-3-(3-methyl-2-butenyl)oxiranyl]-1-oxaspiro[2,5]oct-6-yl(chloroacetyl)carbamate,acetyldinanaline,5-amino-1-[[3,5-dichloro-4-(4-chlorobenzoyl)phenyl]methyl]-1H-1,2,3-triazole-4-carboxamide,CM 101, squalamine, combretastatin, RPI4610, NX31838, sulfatedmannopentaose phosphate,7,7-(carbonyl-bis[imino-N-methyl-4,2-pyrrolocarbonylimino[N-methyl-4,2-pyrrole]-carbonylimino]-bis-(1,3-naphthalenedisulfonate), and 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone(SU5416).

As used above, “integrin blockers” refers to compounds which selectivelyantagonize, inhibit or counteract binding of a physiological ligand tothe α_(v)β₃ integrin, to compounds which selectively antagonize, inhibitor counteract binding of a physiological ligand to the αvβ5 integrin, tocompounds which antagonize, inhibit or counteract binding of aphysiological ligand to both the α_(v)β₃ integrin and the α_(v)β₅integrin, and to compounds which antagonize, inhibit or counteract theactivity of the particular integrin(s) expressed on capillaryendothelial cells. The term also refers to antagonists of the α_(v)β₆,α_(v)β₈, α₁β₁, α₂β₁, α₅β₁, α₆β₁ and α₆β₄ integrins. The term also refersto antagonists of any combination of α_(v)β₃, α_(v)β₅, α_(v)β₆, α_(v)β₈,α₁β₁, α₂β₁, α₅β₁, α₆β₁ and α₆β₄ integrins.

Some specific examples of tyrosine kinase inhibitors includeN-(trifluoromethylphenyl)-5-methylisoxazol-4-carboxamide,3-[(2,4-dimethylpyrrol-5-yl)methylidenyl)indolin-2-one,17-(allylamino)-17-demethoxygeldanamycin,4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-[3-(4-morpholinyl)propoxyl]quinazoline,N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine,BIBX1382,2,3,9,10,11,12-hexahydro-10-(hydroxymethyl)-10-hydroxy-9-methyl-9,12-epoxy-1H-diindolo[1,2,3-fg:3′,2′,1′-k1]pyrrolo[3,4-i][1,6]benzodiazocin-1-one,SH268, genistein, STI571, CEP2563,4-(3-chlorophenylamino)-5,6-dimethyl-7H-pyrrolo[2,3-d]pyrimidinemethanesulfonate, 4-(3-bromo-4-hydroxyphenyl)amino-6,7-dimethoxyquinazoline,4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, SU6668, STI571A,N-4-chlorophenyl-4-(4-pyridylmethyl)-1-phthalazinamine, and EMD121974.

Combinations with compounds other than anti-cancer compounds are alsoencompassed in the instant methods. For example, combinations of theinstantly claimed compounds with PPAR-γ (i.e., PPAR-gamma) agonists andPPAR-δ (i.e., PPAR-delta) agonists are useful in the treatment ofcertain malignancies. PPAR-γ and PPAR-δ are the nuclear peroxisomeproliferator-activated receptors γ and δ. The expression of PPAR-γ onendothelial cells and its involvement in angiogenesis has been reportedin the literature (see J. Cardiovasc. Pharmacol. 1998; 31:909-913; J.Biol. Chem. 1999;274:9116-9121; Invest. Ophthalmol Vis. Sci. 2000;41:2309-2317). More recently, PPAR-γ agonists have been shown to inhibitthe angiogenic response to VEGF in vitro; both troglitazone androsiglitazone maleate inhibit the development of retinalneovascularization in mice. (Arch. Ophthamol. 2001; 119:709-717).Examples of PPAR-γ agonists and PPAR-γ/α agonists include, but are notlimited to, thiazolidinediones (such as DRF2725, CS-011, troglitazone,rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate,GW2570, SB219994, AR-H039242, JTT-501, MCC-555, GW2331, GW409544,NN2344, KRP297, NP0110, DRF4158, NN622, GI262570, PNU182716, DRF552926,2-[(5,7-dipropyl-3-trifluoromethyl-1,2-benzisoxazol-6-yl)oxy]-2-methylpropionicacid WO 01/60807, and2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylicacid WO 02/026729.

Another embodiment of the instant invention is the use of the presentlydisclosed compounds in combination with gene therapy for the treatmentof cancer. For an overview of genetic strategies to treating cancer seeHall et al (Am. J. Hum. Genet. 61:785-789, 1997) and Kufe et al (CancerMedicine, 5th Ed, pp 876-889, B C Decker, Hamilton 2000). Gene therapycan be used to deliver any tumor suppressing gene. Examples of suchgenes include, but are not limited to, p53, which can be delivered viarecombinant virus-mediated gene transfer (see U.S. Pat. No. 6,069,134,for example), a uPA/uPAR antagonist (“Adenovirus-Mediated Delivery of auPA/uPAR Antagonist Suppresses Angiogenesis-Dependent Tumor Growth andDissemination in Mice,” Gene Therapy, August 1998;5(8):1105-13), andinterferon gamma (J. Immunol. 2000;164:217-222).

The compounds of the instant invention may also be administered incombination with an inhibitor of inherent multidrug resistance (MDR), inparticular MDR associated with high levels of expression of transporterproteins. Such MDR inhibitors include inhibitors of p-glycoprotein(P-gp), such as LY335979, XR9576, OC144-093, R101922, VX853 and PSC833(valspodar).

A compound of the present invention may be employed in conjunction withanti-emetic agents to treat nausea or emesis, including acute, delayed,late-phase, and anticipatory emesis, which may result from the use of acompound of the present invention, alone or with radiation therapy. Forthe prevention or treatment of emesis, a compound of the presentinvention may be used in conjunction with other anti-emetic agents,especially neurokinin-1 receptor antagonists, 5HT3 receptor antagonists,such as ondansetron, granisetron, tropisetron, and zatisetron, GABABreceptor agonists, such as baclofen, a corticosteroid such as Decadron(dexamethasone), Kenalog, Aristocort, Nasalide, Preferid, Benecorten orothers such as disclosed in U.S. Pat. Nos. 2,789,118, 2,990,401,3,048,581, 3,126,375, 3,929,768, 3,996,359, 3,928,326 and 3,749,712, anantidopaminergic, such as the phenothiazines (for exampleprochlorperazine, fluphenazine, thioridazine and mesoridazine),metoclopramide or dronabinol. In another embodiment, conjunctive therapywith an anti-emesis agent selected from a neurokinin-1 receptorantagonist, a 5HT3 receptor antagonist and a corticosteroid is disclosedfor the treatment or prevention of emesis that may result uponadministration of the instant compounds.

Neurokinin-1 receptor antagonists of use in conjunction with thecompounds of the present invention are fully described, for example, inU.S. Pat. Nos. 5,162,339, 5,232,929, 5,242,930, 5,373,003, 5,387,595,5,459,270, 5,494,926, 5,496,833, 5,637,699, 5,719,147; European PatentPublication Nos. EP 0 360 390, 0 394 989, 0 428 434, 0 429 366, 0 430771, 0 436 334, 0 443 132, 0 482 539, 0 498 069, 0 499 313, 0 512 901, 0512 902, 0 514 273, 0 514 274, 0 514 275, 0 514 276, 0 515 681, 0 517589, 0 520 555, 0 522 808, 0 528 495, 0 532 456, 0 533 280, 0 536 817, 0545 478, 0 558 156, 0 577 394, 0 585 913,0 590 152, 0 599 538, 0 610793, 0 634 402, 0 686 629, 0 693 489, 0 694 535, 0 699 655, 0 699 674, 0707 006, 0 708 101, 0 709 375, 0 709 376, 0 714 891, 0 723 959, 0 733632 and 0 776 893; PCT International Patent Publication Nos. WO90/05525, 90/05729, 91/09844, 91/18899, 92/01688, 92/06079, 92/12151,92/15585, 92/17449, 92/20661, 92/20676, 92/21677, 92/22569, 93/00330,93/00331, 93/01159, 93/01165, 93/01169, 93/01170, 93/06099, 93/09116,93/10073, 93/14084, 93/14113, 93/18023, 93/19064, 93/21155, 93/21181,93/23380, 93/24465, 94/00440, 94/01402, 94/02461, 94/02595, 94/03429,94/03445, 94/04494, 94/04496, 94/05625, 94/07843, 94/08997, 94/10165,94/10167, 94/10168, 94/10170, 94/11368, 94/13639, 94/13663, 94/14767,94/15903, 94/19320, 94/19323, 94/20500, 94/26735, 94/26740, 94/29309,95/02595, 95/04040, 95/04042, 95/06645, 95/07886, 95/07908, 95/08549,95/11880, 95/14017, 95/15311, 95/16679, 95/17382, 95/18124, 95/18129,95/19344, 95/20575, 95/21819, 95/22525, 95/23798, 95/26338, 95/28418,95/30674, 95/30687, 95/33744, 96/05181, 96/05193, 96/05203, 96/06094,96/07649, 96/10562, 96/16939, 96/18643, 96/20197, 96/21661, 96/29304,96/29317, 96/29326, 96/29328, 96/31214, 96/32385, 96/37489, 97/01553,97/01554, 97/03066, 97/08144, 97/14671, 97/17362, 97/18206, 97/19084,97/19942 and 97/21702; and in British Patent Publication Nos. 2 266 529,2 268 931, 2 269 170, 2 269 590, 2 271 774, 2 292 144, 2 293 168, 2 293169, and 2 302 689. The preparation of such compounds is fully describedin the aforementioned patents and publications, which are incorporatedherein by reference.

In an embodiment, the neurokinin-1 receptor antagonist for use inconjunction with the compounds of the present invention is selectedfrom:2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine,or a pharmaceutically acceptable salt thereof, which is described inU.S. Pat. No. 5,719,147.

A compound of the instant invention may also be administered with anagent useful in the treatment of anemia. Such an anemia treatment agentis, for example, a continuous eythropoiesis receptor activator (such asepoetin alfa).

A compound of the instant invention may also be administered with anagent useful in the treatment of neutropenia. Such a neutropeniatreatment agent is, for example, a hematopoietic growth factor whichregulates the production and function of neutrophils such as a humangranulocyte colony stimulating factor, (G-CSF). Examples of a G-CSFinclude filgrastim.

A compound of the instant invention may also be administered with animmunologic-enhancing drug, such as levamisole, isoprinosine andZadaxin.

A compound of the instant invention may also be useful for treating orpreventing cancer, including bone cancer, in combination withbisphosphonates (understood to include bisphosphonates, diphosphonates,bisphosphonic acids and diphosphonic acids). Examples of bisphosphonatesinclude but are not limited to: etidronate (Didronel), pamidronate(Aredia), alendronate (Fosamax), risedronate (Actonel), zoledronate(Zometa), ibandronate (Boniva), incadronate or cimadronate, clodronate,EB-1053, minodronate, neridronate, piridronate and tiludronate includingany and all pharmaceutically acceptable salts, derivatives, hydrates andmixtures thereof.

Thus, the scope of the instant invention encompasses the use of theinstantly claimed compounds in combination with a second compoundselected from: an estrogen receptor modulator, an androgen receptormodulator, a retinoid receptor modulator, a cytotoxic/cytostatic agent,an antiproliferative agent, a prenyl-protein transferase inhibitor, anHMG-CoA reductase inhibitor, an HIV protease inhibitor, a reversetranscriptase inhibitor, an angiogenesis inhibitor, PPAR-γ agonists,PPAR-δ agonists, an inhibitor of inherent multidrug resistance, ananti-emetic agent, an agent useful in the treatment of anemia, an agentuseful in the treatment of neutropenia, an immunologic-enhancing drug,an inhibitor of cell proliferation and survival signaling, abisphosphonate and an agent that interferes with a cell cyclecheckpoint.

The term “administration” and variants thereof (e.g., “administering” acompound) in reference to a compound of the invention means introducingthe compound or a prodrug of the compound into the system of the animalin need of treatment. When a compound of the invention or prodrugthereof is provided in combination with one or more other active agents(e.g., a cytotoxic agent, etc.), “administration” and its variants areeach understood to include concurrent and sequential introduction of thecompound or prodrug thereof and other agents.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

The term “therapeutically effective amount” as used herein means thatamount of active compound or pharmaceutical agent that elicits thebiological or medicinal response in a tissue, system, animal or humanthat is being sought by a researcher, veterinarian, medical doctor orother clinician.

The term “treating cancer” or “treatment of cancer” refers toadministration to a mammal afflicted with a cancerous condition andrefers to an effect that alleviates the cancerous condition by killingthe cancerous cells, but also to an effect that results in theinhibition of growth and/or metastasis of the cancer.

In an embodiment, the angiogenesis inhibitor to be used as the secondcompound is selected from a tyrosine kinase inhibitor, an inhibitor ofepidermal-derived growth factor, an inhibitor of fibroblast-derivedgrowth factor, an inhibitor of platelet derived growth factor, an MMP(matrix metalloprotease) inhibitor, an integrin blocker, interferon-α,interleukin-12, pentosan polysulfate, a cyclooxygenase inhibitor,carboxyamidotriazole, combretastatin A-4, squalamine,6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin,troponin-1, or an antibody to VEGF. In an embodiment, the estrogenreceptor modulator is tamoxifen or raloxifene.

Also included in the scope of the claims is a method of treating cancerthat comprises administering a therapeutically effective amount of acompound of Formula A in combination with radiation therapy and/or incombination with a second compound selected from: an estrogen receptormodulator, an androgen receptor modulator, a retinoid receptormodulator, a cytotoxiccytostatic agent, an antiproliferative agent, aprenyl-protein transferase inhibitor, an HMG-CoA reductase inhibitor, anHIV protease inhibitor, a reverse transcriptase inhibitor, anangiogenesis inhibitor, PPAR-γ agonists, PPAR-δ agonists, an inhibitorof inherent multidrug resistance, an anti-emetic agent, an agent usefulin the treatment of anemia, an agent useful in the treatment ofneutropenia, an immunologic-enhancing drug, an inhibitor of cellproliferation and survival signaling, a bisphosphonate and an agent thatinterferes with a cell cycle checkpoint.

And yet another embodiment of the invention is a method of treatingcancer that comprises administering a therapeutically effective amountof a compound of Formula A in combination with paclitaxel ortrastuzumab.

The invention further encompasses a method of treating or preventingcancer that comprises administering a therapeutically effective amountof a compound of Formula A in combination with a COX-2 inhibitor.

The instant invention also includes a pharmaceutical composition usefulfor treating or preventing cancer that comprises a therapeuticallyeffective amount of a compound of Formula A and a second compoundselected from: an estrogen receptor modulator, an androgen receptormodulator, a retinoid receptor modulator, a cytotoxic/cytostatic agent,an antiproliferative agent, a prenyl-protein transferase inhibitor, anHMG-CoA reductase inhibitor, an HIV protease inhibitor, a reversetranscriptase inhibitor, an angiogenesis inhibitor, a PPAR-γ agonist, aPPAR-δ agonist, an inhibitor of cell proliferation and survivalsignaling, a bisphosphonate, and an agent that interferes with a cellcycle checkpoint.

All patents, publications and pending patent applications identified arehereby incorporated by reference.

Abbreviations used in the description of the chemistry and in theExamples that follow are: AEBSF (p-aminoethylbenzenesulfonyl fluoride);Atm (atmosphere); BSA (bovine serum albumin); Boc (tert-butoxycarbonyl);BuLi (n-Butyl lithium); CDCl₃ (chloroform-d); CuI (copper iodide); CuSO₄(copper sulfate); DCE (dichloroethane); DCM (dichloromethane); DEAD(diethyl azodicarboxylate); DMF (N,N-dimethylformamide); DMSO (dimethylsulfoxide); DTT (dithiothreitol); EDTA (ethylene-diamine-tetra-aceticacid); EGTA (ethylene-glycol-tetra-acetic acid); EtOAc (ethyl acetate);EtOH (ethanol); HOAc (acetic acid); HPLC (high-performance liquidchromatography); HRMS (high resolution mass spectrum); LCMS (liquidchromatograph-mass spectrometer); LHMDS (lithiumbis(trimethylsilyl)amide); LRMS (low resolution mass spectrum); Me(methyl); MeOH (methanol); MP- B(CN)H₃ (Macroporous cyanoborohydride);NaHCO₃ (sodium bicarbonate); Na₂SO₄ (sodium sulfate); Na(OAc)₃BH (sodiumtriacetoxyborohydride); NH₄OAc (ammonium acetate); NBS(N-bromosuccinamide); NMR (nuclear magnetic resonance); PBS (phosphatebuffered saline); PCR (polymerase chain reaction); Pd(dppf)([1,1′-bis(diphenylphosphino)ferrocene]palladium); Pd(Ph₃)₄(palladium(0) tetrakis-triphenylphosphine); POCl₃ (phosphorousoxychloride); PS-DIEA (polystyrene diisopropylethylamine); PS-PPh₃(polystyrene-triphenyl phosphine); TBAF (tetrabutylammonium fluoride);THF (tetrahydrofuran); TFA (trifluoroacteic acid); and TMSCH₂N₂(trimethylsilyldiazomethane).

The compounds of this invention may be prepared by employing reactionsas shown in the following Reaction Schemes, in addition to otherstandard manipulations that are known in the literature or exemplifiedin the experimental procedures. The illustrative Reaction Schemes below,therefore, are not limited by the compounds listed or by any particularsubstituents employed for illustrative purposes. Substituent numberingas shown in the Reaction Schemes does not necessarily correlate to thatused in the claims and often, for clarity, a single substituent is shownattached to the compound where multiple substituents are allowed underthe definitions of Formula A hereinabove.

Reactions that may be used to generate the compounds of this inventionare prepared by employing reactions as shown in the Reaction SchemesI-X, in addition to other standard manipulations such as esterhydrolysis, cleavage of protecting groups, etc., as may be known in theliterature or exemplified in the experimental procedures.

These reactions may be employed in a linear sequence to provide thecompounds of the invention or they may be used to synthesize fragmentswhich are subsequently joined by the alkylation reactions described inthe Reaction Schemes.

Synopsis of Reaction Schemes

The following Reaction Schemes, Reaction Schemes I-VII, provide usefuldetails for preparing the bicyclic moieties of the instant compounds.Aryl groups may be substituted with heterocylic moieties followingReactions Schemes VIII-IX.

The requisite intermediates are in some cases commercially available, orcan be prepared according to literature procedures. As illustrated inReaction Scheme I, a suitably substituted phenylacetylide may be reactedwith copper iodide to form the corresponding copper acetylide I-1 (seefor example, Sonogashira, K.; Toda, Y.; Hagihara, N. Tetrahedron Lett.1975, 4467). Intermediate I may then react with a suitably substitutedelectrophilic moiety to provide the asymmetrically substituted I-2.Reaction with NBS followed by hydrolysis provides I-3 (see for example,Yusybov, M. S.; Filimonov, V. D.; Synthesis 1991, 2, 131). A variety ofsubstituted and unsubstituted aryls and heterocyclyls may also beobtained commercially.

Reaction Scheme II illustrates the preparation of the compounds,starting with a suitably substituted II-1. This intermediate can bereacted with a suitably substituted amine to provide intermediate II-2,which can be reacted with a suitable aryl or heteroaryl diamine toprovide a regioisomeric mixture of the instant compounds, which canusually be separated chromatagraphically.

Reaction Scheme III illustrates the synthesis of a compound wherein “y”is CH and “z” is N.

Reaction Scheme IV illustrates the preparation of compounds, startingwith a suitably substituted 4-amino-3-nitrobenzonitrile IV-I. Thisintermediate can then subjected to a microwave promoted [3+2]cycloaddition reaction to afford tetrazole IV-II. Alkylation of theacidic tetrazole with an electrophile, such as methyl iodide, delivers a2-methyl(IV-III)/1-methyl(IV-IV) mixture of alkylated tetrazoles whichare separable by column chromatography. Ra—Ni hydrogenation of IV-IIIdelivers the diamine IV-V. Further synthesis is as described in theReaction Schemes above.

Reaction Scheme V is illustrative for the synthesis of compounds.Friedlander cyclocondensation of a suitably substituted aryl orheteroaryl aminoaldehyde with a suitably substituted ketone givesintermediates of formula V-1. Conversion of the carboxylic acidfunctionality to an aldehyde functionality gives intermediate V-2 and isaccomplished by methods well known to someone skilled in the art.Reductive alkylation with a suitably substituted amine providescompounds of formula V-3.

Reaction Scheme VI illustrates an alternate synthesis of intermediateV-2.

Reaction Scheme VII illustrates the synthesis of the compounds, startingwith ketone VII-1 which is prepared according to literature (Renault,O.; Dallemagne, P.; and Rault, S. Org. Prep. Proced. Int., 1999, 31,324). Condensation of VII-1 with N,N-dimethylformamide dimethylacetalgives keto-enamine VII-2, which cyclizes with 2-cyanoacetamide to affordpyridone VII-3. Treatment of VII-3 with phosphorus oxychloride produceschloropyridin VII-4. Radical bromination followed by displacement withsuitably substituted amines generates amines VII-5. Subsequent reactionof chloronicotinonitriles VII-5 with various bisnucleophiles providesthe cyclized structures VII-6.

Reactions used to generate the compounds of this invention are preparedby employing reactions as shown in the Reaction Schemes VIII and IX, inaddition to other standard manipulations such as ester hydrolysis,cleavage of protecting groups, etc., as may be known in the literatureor exemplified in the experimental procedures.

Further Synopsis of Reaction Schemes

Reaction Scheme VIII illustrates the preparation of the1,6-naphthyridin-6(5H)-one compounds. Synthesis starts with acommercially available carboxylic acid (VIII-1), which is converted toWeinreb amide (VIII-2). Reaction of amide with an aryl lithium reagent,generated through lithium-halogen exchange reaction of an aryl bromidewith n-butyl lithium, provides the ketone (VIII-3) Condensation of thissubstituted ketone with 4-aminonicotinaldehyde in the presence of abase, for example, sodium hydroxide or sodium methoxide, gives the1,6-naphthyridine (VIII-4). The R substituent on the phenyl can be afunctional group, such as, a (silyl)protected hydroxymethyl, the maskedaldehyde (i.e. acetal), or a carboxylic acid. This material can beconverted to the requisite aldehyde VIII-4. When the R group ishydroxymethyl, oxidation with a reagent such as activated manganesedioxide gives the aldehyde VIII-4. When R is an acetal, mild acidhydrolysis gives the aldehyde. Reduction of the carboxylic acid groupvia reduction of a mixed anhydride with a borohydride also gives thealdehyde VIII-4. This aldehyde can then undergo reductive amination witha diverse array of amines, for example, a 4-subtituted piperidine, and aborohydride to provide 1,6-naphthyridin-5(H)-ones (VIII-5). Treatment of(VIII-5) with pyridinium hydrochloride at 150° C. furnishes the finalproduct (VIII-6).

Reaction Scheme IX illustrates an alternate strategy of introducingheterocycles at the C3-position. It starts with commercially availabledichloro benzopyrazine, which can be coupled to an aryl boronic acid inthe presence of a palladium catalyst (Suzuki reaction). Then a repeat ofthis reaction with another heterocyclic boronic acid provides the finalproduct, under the same reaction conditions.

Reaction Scheme X illustrates an alternative preparation of the1,6-naphthyridin-5(6H)-one compounds. In this Scheme, Weinreb amideVIII-2 is reacted with an aryl lithium reagent, generated from arylbromide X-1, in which R is a protected or masked amino methyl derivativesuch as 1-(1-azido-1-methylethyl), to give ketone X-2. Condensation ofthis ketone with tert-butyl(2-chloro-3-formylpyridin-4-yl)carbamate inthe presence of sodium methoxide, gives the 1,6-naphthyridine X-3.Deprotection or unmasking of the amine, in this case by palladiumcatalyzed reduction with hydrogen provides amine X-4. Treatment of X-4with hydrochloric acid gives 1,6-naphthyridin-5(6H)-one X-5.

EXAMPLES

Examples and schemes provided are intended to assist in a furtherunderstanding of the invention. Particular materials employed, speciesand conditions are intended to be further illustrative of the inventionand not limitative of the reasonable scope thereof.

5-Methoxy-2(4-{[4-5-pyridin-2-yl-4H-1,2,4-triazol)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridine(1-7) and2-(4-{[4-(5-Pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one(1-8) N-Methoxy-N-methyl-2-(2-thienyl)acetamide (1-1)

A 25 mL RB flask with a stirring bar was charged with 2-thiopheneaceticacid (2.84 g, 2 mmol), followed by anhydrous DMF (10 mL).1,1′-Carbonyldiimidazole (3.24 g, 2 mmol) was added in one portion,giving rise to significant gas evolution. The mixture was warmed to 40°C. After 30 min, N,O-dimethylhydroxyamine hydrochloride (2.14 g, 2.2mmol) was introduced in one portion. The mixture was allowed to stir at23° C. for 30 min. before being diluted with sat. ammonium chloridesolution (100 mL) and extracted twice with 1:1 ethyl acetate/hexanes(100 mL). The crude mixture was purified by column chromatography oversilica gel (0-25% ethyl acetate/hexanes) to giveN-methoxy-N-methyl-2-(2-thienyl)acetamide (1-1). ¹H NMR (CDCl₃, ppm) δ7.20 (m, 1H), 6.96 (m, 2H), 3.91 (s, 2H), 3.70 (s, 3H), 3.25 (s, 3H).

1-[4-(1,3-Dioxolan-2-yl)phenyl]-2-(2-thienyl)ethanone (1-2)

A 25 ml RB flask with a stirring bar was charged with1-bromo-4-(1,3-dioxolan-2-yl)benzene (243 mg, 1.0 mmol), followed byanhydrous THF (5 mL). The mixture was cooled to −78° C. andn-butyllithium (687 μL of 1.6 M solution in hexanes) was added dropwiseover 2 min. After stirring for 15 min. at −78° C.,N-methoxy-N-methyl-2-(2-thienyl)acetamide (1-1) (185 mg, 1 mmol) wasadded in one portion. After another 30 min at −78° C., the mixture wasquenched with sat. ammonium chloride (20 mL), extracted twice with ethylacetate (10 mL). The combined organic extracts were dried over sodiumsulfate, concentrated, and purified via column chromatography oversilica gel (0-50% ethyl acetate/hexanes) to give1-[4-(1,3-dioxolan-2-yl)phenyl]-2-92-thienyl)ethanone (1-2). ¹H NMR(CDCl₃, ppm) δ: 7.98 (d, J=8.25 Hz, 2H), 7.45 (d, J=8.25 Hz, 2H), 7.15(d, J=5.15 Hz, 1H), 6.86 (m, 1H), 6.83 (m, 1H), 5.77 (s, 1H), 4.38 (s,2H), 4.01 (m, 4H).

tert-Butyl (2-chloro-3-formylpyridin-4-yl)carbamate (1-3)

To 4-amino-2-chloropyridine (30.0 g, 243 mmol) in CH₂Cl₂ (500 mL) wasadded triethylamine (38 mL), di-tert-butyl-dicarbonate (58.0 g), andDMAP (10.0 g). The reaction mixture was stirred overnight, concentratedin vacuo, and chromatographed on silica gel with 1:1 hexanes/EtOAc. Theresulting solid was triturated with CH₂Cl_(2/)EtOAc (1:1) to giveN-BOC-4-amino-2-chloropyridine. ¹H NMR: (500 MHz, CDCl₃) δ 8.19 (d, 1H),7.53 (d, 1H), 7.25 (br s, 1H), 7.20 (dd, 1H), 5.80 (s, 1H), 1.52 (s,9H).

To N-Boc-4-amino-2-chloropyridine (14.4 g, 63 mmol) in THF (200 mL) at−78° C. was added a 1.7M solution of tert-butyllithium (100 mL) dropwiseover 10 minutes and the solution stirred for 2 h. DMF (15 mL) was thenadded and the reaction allowed to reach ambient temperature over 3 h.The reaction was carefully quenched with saturated NH₄Cl, extracted withEtOAc, and the organic layer washed with H₂O and brine, dried overMgSO₄, filtered, concentrated in vacuo, and purified by silica gelchromatography (10-25% EtOAc/hexanes) to givetert-butyl(2-chloro-3-formylpyridin-4-yl)carbamate (1-3) as a whitesolid. ¹H NMR: (500 MHz, CDCl₃) δ 10.98 (br s, 1H), 10.49 (s, 1H), 8.35(d, 1H), 8.28 (d, 1H), 1.52 (s, 9H).

4-[5-Methoxy-3-(2-thienyl)-1,6-naphthyridin-2-yl]benzaldehyde (1-5)

A 25-ml RB flask with a stirring bar was charged with1-[4-(1,3-dioxolan-2-yl)phenyl]-2-(2-thienyl)ethanone (1-2) (80 mg, 0.29mmol), tert-butyl(2-chloro-3-formylpyridin-4-yl)carbamate (1-3) (90 mg,0.35 mmol) and anhydrous methanol (2 mL). Then sodium methoxide (25% w/wsolution in methanol) was added in one portion and the mixture washeated to 65° C. for 4 h, giving rise to a dark orange suspension.Solvent was removed via rotavap and the mixture was acidified with 1Mhydrochloric acid (10 mL), extracted twice with ethyl acetate (10 mL).The crude mixture was purified by column chromatography on silica gel(0-50% ethyl acetate/hexanes) to give2-[4-(1,3-dioxolan-2-yl)phenyl]-5-methoxy-3-phenyl-1,6-naphthyridine(1-4).

The 2-[4-(1,3-dioxolan-2-yl)phenyl]-5-methoxy-3-phenyl-1,6-naphthyridine(1-4) was dissolved in a mixture of THF (4 mL) and 1M HCl (4 mL)solution, and stirred at 23° C. for 2 h until no starting material waspresent as determined by TLC. The reaction mixture was diluted with sat.sodium bicarbonate solution (10 mL) and extracted twice with ethylacetate (10 mL). Concentration of the combined organic extracts gave4-[5-methoxy-3-(2-thienyl)-1,6-naphthyridin-2-yl]benzaldehyde (1-5). ¹HNMR (CDCl₃, ppm) δ: 9.93 (s, 1H), 8.54 (s,1H), 8.12 (d, J=6.0 Hz, 1H),7.75 (d, J=8.2 Hz, 2H), 7.40 (d, J=8.2 Hz, 2H), 7.19 (d, J=5.0 Hz, 1H),7.03 (d, J=3.5 Hz, 1H), 6.82 (d, J=3.6 Hz, 1H), 6.69 (s, 1H), 4.13 (s,3H).

5-Methoxy-2(4-{[4-5-pyridin-2-yl-4H-1,2,4-triazol)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridine(1-7)

A 25 ml RB flask with a stirring bar was charged with4-[5-methoxy-3-(2-thienyl)-1,6-naphthyridin-2-yl]benzaldehyde (1-5) (62mg, 0.18 mmol), followed by2-(3-piperidin-4-yl-1H-1,2,4-triazol-5-yl)pyridine dihydrochloride (1-6)(82 mg, 0.36 mmol), anhydrous DMF (1.0 mL), triethylamine (72 mg, 0.716mmol), and acetic acid (107 mg, 1.79 mmol). Then sodiumtriacetoxyborohydride (76 mg, 0.36 mmol) was added in one portion, andthe resulting pale yellow solution was stirred at 23° C. overnight. Themixture was diluted with sat. ammonium chloride solution (10 mL),extracted twice with ethyl acetate (10 mL). The crude mixture waspurified by column chromatography on silica gel (0-30%methanol/dichloromethane) to give5-methoxy-2(4-{[4-5-pyridin-2-yl-4H-1,2,4-triazol)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridine(1-7). ¹H NMR (CDCl₃, ppm) δ: 8.54 (bs, 1H), 8.51 (s,1H), 8.10 (d, J=6.0Hz, 1H), 8.01 (d, J=7.8 Hz, 1H), 7.68 (t, J=7.7 Hz, 1H), 7.42 (d, J=8.0Hz, 2H), 7.37 (d, J=6.9 Hz, 1H), 7.29 (d, J=8.0 Hz, 2H), 7.25 (t, J=3.4Hz, 1H), 7.19 (d, J=5.1 Hz, 1H), 7.15 (s, 1H), 6.81 (t, J=3.6 Hz, 1H),6.68 (d, J=3.1 Hz, 1H), 4.05 (s, 3H), 3.98 (s, 2H), 3.35 (s, 1H), 3.25(bs, 2H), 3.02 (bs, 2H), 2.75 (bs, 2H), 2.19 (bs, 2H).

2-(4-{[4-(5-Pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one(1-8)

A 10 ml RB flask with a stirring bar was charged with5-methoxy-2(4-{[4-5-pyridin-2-yl-4H-1,2,4-triazol)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridine(1-7) (40 mg, 0.7 mmol) and pyridinium hydrochloride (738 mg, 6.4 mmol).The mixture was heated to 150° C. for 10 min. until the mixture meltedinto a yellow solution. The mixture solidified again upon cooling, wasneutralized with sat. sodium bicarbonate solution (10 mL), and extractedtwice with ethyl acetate (10 mL). The combined organic extracts wereconcentrated and purified by column chromatography over silica gel(0-30% methanol/dichloromethane) to furnish2-(4-{[4-(5-pyridin-2-yl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one(1-8). ¹H NMR (CDCl₃, ppm) δ: 8.69 (s, 1H), 8.51 (s,1H), 8.40 (d, J=6.0Hz, 1H), 8.03 (d, J=7.8 Hz, 1H), 7.76 (t, J=7.7 Hz, 1H), 7.39 (d, J=8.0Hz, 2H), 7.27 (m, 4H), 7.22 (d, J=4.4 Hz, 1H), 6.78 (d, J=5.1 Hz, 2H),3.55 (s, 2H), 2.95 (d, J=11.6 Hz, 2H), 2.83 (bs, 1H), 2.14 (m, 2H), 1.97(m, 2H), 1.87 (m, 2H).

The following compounds in Table 1 were prepared in a similar manner asshown in Scheme 1 and as shown in the Reaction Schemes:

TABLE 1 LRMS Cmp Structure Name m/z (M + H) 1-9

1-{4-[4-(3-pyrimidin-5- ylquinoxalin-2- yl)benzyl]cyclohexyl}-1,3-dihydro-2H- benimidazol-2-one 513.2 1-10

3-[3-(4-{[4-2-oxo-2,3- dihydro-1H-benimidazol- 1-yl)cyclohexyl]methyl}phenyl)quinoxalin-2- yl]thiophene-2- carbaldehyde 545.2 1-11

1-(4-{4-[3-(1 H-pyrazol-5- yl)quinoxalin-2- yl]benzyl}cyclohexyl)-1,3-dihydro-2H- benzimidazol-2-one 502.2

2-[4-(1-amino-1-methylethyl)phenyl]-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one(2-7) 2-(4-bromophenyl)propan-2-ol (2-2)

Methylmagnesium bromide solution (1.4M in 75:25 toluene:THF, 20 mL, 28mmol) was added slowly to ethyl 4-bromobenzoate (2-1, 2.52 g, 11.0 mmol)in THF (10 mL) at −30° C. After 2 hr, the mixture was quenched withammonium chloride and extracted with ether. The organic layer was washedwith 1:1 brine:water, dried over magnesium sulfate, filtered andconcentrated to give the title compound as a pale yellow oil (J. Am.Chem. Soc. 1971, 93, 6877).

1-(1-azido-1-methylethyl)-4-bromobenzene (2-3)

A solution of TFA (4.4 mL, 45.0 mmol) in chloroform (10 mL) was addedslowly to a mixture of 2-(4-bromophenyl)propan-2-ol (2-2, 2.33 g, 10.8mmol) and sodium azide (1.42 g, 21.8 mmol) in chloroform (10 mL) cooledto −5° C., maintaining the temperature below 0° C. The cooling bath wasremoved and the mixture was stirred overnight at room temperature.Concentrated ammonium hydroxide was added until basic (moist pH paper).The organic layer was washed with 1:1 brine:water, dried over magnesiumsulfate, filtered and concentrated to give the title compound (2.51 g)as a pale yellow oil. LRMS m/z (M-N₂) Calcd: 197.0,. Found 197.1.

1-[4-(1-azido-1-methylethyl)phenyl]-2-(2-thienyl)ethanone (2-4)

nBuLi (1.6M in hexane, 2.88 mL, 4.60 mmol) was added dropwise to asolution of 1-(1-azido-1-methylethyl)-4-bromobenzene (2-3, 1.03 g, 4.28mmol) in THF (20 mL) at −78° C. After 15 min a solution ofN-methoxy-N-methyl-2-thien-2-ylacetamide (1-1, 0.78 g, 4.18 mmol) in THF(1 mL) was added. After an additional 30 min at −78° C., the reactionwas quenched with saturated ammonium chloride solution and extractedwith ethyl acetate, dried over magnesium sulfate, filtered, andconcentrated to give an oil. The crude product was purified viaautomated flash column chromatography on silica gel (5% CH₂Cl₂, 0-10%EtOAc/hexanes over 30 min) to give the title compounds as a yellow oil.LRMS m/z (M+1) Calcd: 286.1,. Found 286.2.

2-[4-(1-azido-1-methylethyl)phenyl]-5-methoxy-3-(2-thienyl)-1,6-naphthyridine(2-5)

Sodium methoxide (25 weight % in methanol, 0.23 mL) was added to astirred mixture of1-[4-(1-azido-1-methylethyl)phenyl]-2-(2-thienyl)ethanone (2-4, 95mg,0.33mmol) and tert-butyl(2-chloro-3-formylpyridin-4-yl)carbamate (1-3,100 mg, 0.39 mmol) in methanol (2 mL), then heated to 65° C. for 2 h.The mixture was cooled to rt, diluted with EtOAc, and acidified with 1NHCl. The aqueous was extracted with ethyl acetate and the combinedorganics were washed with 1:1 brine:water, dried over magnesium sulfate,filtered, and concentrated to give a dark semi-solid. The crude productwas purified via automated silica gel chromatography (0-45% EtOAc inhexane with 5% CH₂Cl₂ over 30 min) to give the title compound as anamber oil. LRMS m/z (M+1) Calcd: 402.2,. Found 402.2.

2-{4-[5-methoxy-3-(2-thienyl)-1,6-naphthyridin-2-yl]phenyl}propan-2-amine(2-6)

A mixture of2-[4-(1-azido-1-methylethyl)phenyl]-5-methoxy-3-(2-thienyl)-1,6-naphthyridine(2-5, 78 mg, 0.194 mmol) and 10% Pd/C (9 mg) was stirred under 1atmhydrogen in EtOH (5 mL) for 2 h. The mixture was filtered through celiteand concentrated to give the title compound as a clear oil. LRMS m/z(M+1) Calcd: 376.1,. Found 376.2.

2-[4-(1-amino-1-methylethyl)phenyl]-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one(2-7)

To a solution of2-{4-[5-methoxy-3-(2-thienyl)-1,6-naphthyridin-2-yl]phenyl}propan-2-amine(2-6, 66 mg, 0.176 mmol) in THF (10 mL) was added concentrated HCl (1mL) and stirred for 5 h at rt. The volatiles were evaporated in vacuoand the residue was partitioned between saturated sodium bicarbonatesolution and ethyl acetate. The organic layer was dried over magnesiumsulfate, filtered, and concentrated to give the title compound as anoff-white solid. LRMS m/z (M+1) Calcd: 362.1,. Found 362.2.

Example 1

Cloning of the Human Akt Isoforms and ΔPH-Akt1

The pS2neo vector (deposited in the ATCC on Apr. 3, 2001 as ATCCPTA-3253) was prepared as follows: The pRmHA3 vector (prepared asdescribed in Nucl. Acid Res. 16:1043-1061 (1988)) was cut with BglII anda 2734 bp fragment was isolated. The pUChsneo vector (prepared asdescribed in EMBO J. 4:167-171 (1985)) was also cut with BglII and a4029 bp band was isolated. These two isolated fragments were ligatedtogether to generate a vector termed pS2neo-1. This plasmid contains apolylinker between a metallothionine promoter and an alcoholdehydrogenase poly A addition site. It also has a neo resistance genedriven by a heat shock promoter. The pS2neo-1 vector was cut with Psp5IIand BsiWI. Two complementary oligonucleotides were synthesized and thenannealed (CTGCGGCCGC (SEQ.ID.NO.: 1) and GTACGCGGCCGCAG (SEQ.ID.NO.:2)). The cut pS2neo-1 and the annealed oligonucleotides were ligatedtogether to generate a second vector, pS2neo. Added in this conversionwas a NotI site to aid in the linearization prior to transfection intoS2 cells.

Human Akt1 gene was amplified by PCR (Clontech) out of a human spleencDNA (Clontech) using the 5′ primer:5′CGCGAATTCAGATCTACCATGAGCGACGTGGCTATTGTG 3′ (SEQ.ID.NO.: 3), and the 3′primer: 5′CGCTCTAGAGGATCCTCAGGCCGTGCTGCTGGC3′ (SEQ.ID.NO.: 4). The 5′primer included an EcoRI and BglII site. The 3′ primer included an XbaIand BamHI site for cloning purposes. The resultant PCR product wassubcloned into pGEM3Z (Promega) as an EcoRI/Xba I fragment. Forexpression/purification purposes, a middle T tag was added to the 5′ endof the full length Akt1 gene using the PCR primer:5′GTACGATGCTGAACGATATCTTCG 3′ (SEQ.ID.NO.: 5). The resulting PCR productencompassed a 5′ KpnI site and a 3′ BamHI site which were used tosubclone the fragment in frame with a biotin tag containing insect cellexpression vector, pS2neo.

For the expression of a pleckstrin homology domain (PH) deleted (Δaa4-129, which includes deletion of a portion of the Akt1 hinge region)version of Akt1, PCR deletion mutagenesis was done using the full lengthAkt1 gene in the pS2neo vector as template. The PCR was carried out in 2steps using overlapping internal primers(5′GAATACATGCCGATGGAAAGCGACGGGGCTGAAGAGATGGAGGTG 3′ (SEQ.ID.NO.: 6), and5′CCCCTCCATCTCTTCAGCCCCGTCGCTTTCCATCGGCATG TATTC 3′ (SEQ.ID.NO.: 7))which encompassed the deletion and 5′ and 3′ flanking primers whichencompassed the KpnI site and middle T tag on the 5′ end. The final PCRproduct was digested with KpnI and SmaI and ligated into the pS2neo fulllength Akt1 KpnI/SmaI cut vector, effectively replacing the 5′ end ofthe clone with the deleted version.

Human Akt3 gene was amplified by PCR of adult brain cDNA (Clontech)using the amino terminal oligo primer: 5′GAATTCAGATCTACCATGAGCGATGTTACCATTGTG 3′ (SEQ.ID.NO.: 8); and the carboxyterminal oligo primer: 5′ TCTAGATCTTATTCTCGTCCACTTGCAGAG 3′(SEQ.ID.NO.:9). These primers included a 5′ EcoRI/BglII site and a 3′ XbaI/BglIIsite for cloning purposes. The resultant PCR product was cloned into theEcoRI and XbaI sites of pGEM4Z (Promega). For expression/purificationpurposes, a middle T tag was added to the 5′ end of the full length Akt3clone using the PCR primer:5′GGTACCATGGAATACATGCCGATGGAAAGCGATGTTACCATTGTGAAG 3′(SEQ.ID.NO.: 10).The resultant PCR product encompassed a 5′ KpnI site which allowed inframe cloning with the biotin tag containing insect cell expressionvector, pS2neo.

Human Akt2 gene was amplified by PCR from human thymus cDNA (Clontech)using the amino terminal oligo primer: 5′AAGCTTAGATCTACCATGAATGAGGTGTCTGTC 3′ (SEQ.ID.NO.: 11); and the carboxyterminal oligo primer: 5′GAATTCGGATCCTCACTCGCGGATGCTGGC 3′ (SEQ.ID.NO.:12). These primers included a 5′ HindIII/BglII site and a 3′ EcoRI/BamHIsite for cloning purposes. The resultant PCR product was subcloned intothe HindIII/EcoRI sites of pGem3Z (Promega). For expression/purificationpurposes, a middle T tag was added to the 5′ end of the full length Akt2using the PCR primer:5′GGTACCATGGAATACATGCCGATGGAAAATGAGGTGTCTGTCATCAAAG 3′ (SEQ.ID.NO.: 13).The resultant PCR product was subcloned into the pS2neo vector asdescribed above.

Example 2

Expression of Human Akt Isoforms and ΔPH-Akt1

The DNA containing the cloned Akt1, Akt2, Akt3 and ΔPH-Akt1 genes in thepS2neo expression vector was purified and used to transfect DrosophilaS2 cells (ATCC) by the calcium phosphate method. Pools of antibiotic(G418, 500 μg/ml) resistant cells were selected. Cell were expanded to a1.0 L volume (˜7.0×10⁶/ml), biotin and CuSO₄ were added to a finalconcentration of 50 μM and 50 mM respectively. Cells were grown for 72 hat 27° C. and harvested by centrifugation. The cell paste was frozen at−70° C. until needed.

Example 3

Purification of Human Akt Isoforms and ΔPH-Akt1

Cell paste from one liter of S2 cells, described in Example 2, was lysedby sonication with 50 mls 1% CHAPS in buffer A: (50 mM Tris pH 7.4, 1 mMEDTA, 1 mM EGTA, 0.2 mM AEBSF, 10 μg/ml benzamidine, 5 μg/ml ofleupeptin, aprotinin and pepstatin each, 10% glycerol and 1 mM DTT). Thesoluble fraction was purified on a Protein G Sepharose fast flow(Pharmacia) column loaded with 9 mg/ml anti-middle T monoclonal antibodyand eluted with 75 μM EYMPME (SEQ.ID.NO.: 14) peptide in buffer Acontaining 25% glycerol. Akt/PKB containing fractions were pooled andthe protein purity evaluated by SDS-PAGE. The purified protein wasquantitated using a standard Bradford protocol. Purified protein wasflash frozen on liquid nitrogen and stored at −70° C.

Akt and Akt pleckstrin homology domain deletions purified from S2 cellsrequired activation. Akt and Akt pleckstrin homology domain deletionswere activated (Alessi et al. Current Biology 7:261-269) in a reactioncontaining 10 nM PDK1 (Upstate Biotechnology, Inc.), lipid vesicles (10μM phosphatidylinositol-3,4,5-trisphosphate—Metreya, Inc, 100 μMphosphatidylcholine and 100 μM phosphatidylserine—Avanti Polar lipids,Inc.) and activation buffer (50 mM Tris pH7.4, 1.0 mM DTT, 0.1 mM EGTA,1.0 μM Microcystin-LR, 0.1 mM ATP, 10 mM MgCl₂, 333 μg/ml BSA and 0.1 mMEDTA). The reaction was incubated at 22° C. for 4 hours. Aliquots wereflash frozen in liquid nitrogen.

Example 4

Akt Kinase Assays

Activated Akt isoforms and pleckstrin homology domain deletionconstructs were assayed utilizing a GSK-derived biotinylated peptidesubstrate. The extent of peptide phosphorylation was determined byHomogeneous Time Resolved Fluorescence (HTRF) using a lanthanidechelate(Lance)-coupled monoclonal antibody specific for thephosphopeptide in combination with a streptavidin-linked allophycocyanin(SA-APC) fluorophore which will bind to the biotin moiety on thepeptide. When the Lance and APC are in proximity (i.e. bound to the samephosphopeptide molecule), a non-radiative energy transfer takes placefrom the Lance to the APC, followed by emission of light from APC at 665nm.

Materials required for the assay:

-   A. Activated Akt isozyme or pleckstrin homology domain deleted    construct-   B. Akt peptide substrate: GSK3α (S21) Peptide #3928    biotin-GGRARTSSFAEPG (SEQ.ID.NO.:15), Macromolecular Resources.-   C. Lance labeled anti-phospho GSK3α monoclonal antibody (Cell    Signaling Technology, clone #27).-   D. SA-APC (Prozyme catalog no. PJ25S lot #896067).-   E. Microfluor®B U Bottom Microtiter Plates (Dynex Technologies,    Catalog no. 7205).-   F. Discovery® HTRF Microplate Analyzer, Packard Instrument Company.-   G. 100 X Protease Inhibitor Cocktail (PIC): 1 mg/ml benzamidine, 0.5    mg/ml pepstatin, 0.5 mg/ml leupeptin, 0.5 mg/ml aprotinin.-   H. 10× Assay Buffer: 500 mM HEPES, pH 7.5, 1% PEG, mM EDTA, 1 mM    EGTA, 1% BSA, 20 mM θ-Glycerol phosphate.-   I. Quench Buffer: 50 mM HEPES pH 7.3, 16.6 mM EDTA, 0.1% BSA, 0.1%    Triton X-100, 0.17 nM Lance labeled monoclonal antibody clone #27,    0.0067 mg/ml SA-APC-   J. ATP/MgCl₂ working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 125    mM KCl, 5% Glycerol, 25 mM MgCl₂, 375 TM ATP-   K. Enzyme working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 5%    Glycerol, active Akt. The final enzyme concentrations were selected    so that the assay was in a linear response range.-   L. Peptide working solution: 1× Assay buffer, 1 mM DTT, 1×PIC, 5%    Glycerol, 2 TM GSK3 biotinylated peptide #3928

The reaction is assembled by adding 16 TL of the ATP/MgCl₂ workingsolution to the appropriate wells of a 96-well microtiter plate.Inhibitor or vehicle (1.0 Tl) is added followed by 10 Tl of peptideworking solution. The reaction is started by adding 13 Tl of the enzymeworking solution and mixing. The reaction is allowed to proceed for 50min and then stopped by the addition of 60 Tl HTRF quench buffer. Thestopped reactions were incubated at room temperature for at least 30 minand then read on the Discovery instrument.

Procedure for Streptavidin Flash Plate Assay:

Step 1:

A 1 μl solution of the test compound in 100% DMSO was added to 20 μl of2× substrate solution (20 uM GSK3 Peptide, 300 μM ATP, 20 mM MgCl₂, 20μCi/ml [γ³³P] ATP, 1× Assay Buffer, 5% glycerol, 1 mM DTT, 1×PIC, 0.1%BSA and 100 mM KCl). Phosphorylation reactions were initiated by adding19 μl of 2× Enzyme solution (6.4 nM active Akt/PKB, 1× Assay Buffer, 5%glycerol, 1 mM DTT, 1×PIC and 0.1% BSA). The reactions were thenincubated at room temperature for 45 minutes.

Step 2:

The reaction was stopped by adding 170 μl of 125 mM EDTA. 200 μl ofstopped reaction was transferred to a Streptavidin Flashplate® PLUS (NENLife Sciences, catalog no. SMP103). The plate was incubated for ≧10minutes at room temperature on a plate shaker. The contents of each wellwas aspirated, and the wells rinsed 2 times with 200 μl TBS per well.The wells were then washed 3 times for 5 minutes with 200 μl TBS perwell with the plates incubated at room temperature on a platform shakerduring wash steps.

The plates were covered with sealing tape and counted using the PackardTopCount with the appropriate settings for counting [³³P] inFlashplates.

Procedure for Streptavidin Filter Plate Assay:

Step 1:

The enzymatic reactions as described in Step 1 of the Streptavidin FlashPlate Assay above were performed.

Step 2:

The reaction was stopped by adding 20 μl of 7.5M GuanidineHydrochloride. 50 μl of the stopped reaction was transferred to theStreptavidin filter plate (SAM²™ Biotin Capture Plate, Promega, catalogno. V7542) and the reaction was incubated on the filter for 1-2 minutesbefore applying vacuum.

The plate was then washed using a vacuum manifold as follows: 1) 4×200μl/well of 2M NaCl; 2) 6×200 μl/well of 2M NaCl with 1% H₃PO₄; 3) 2×200μl/well of diH₂0; and 4) 2×100 μl/well of 95% Ethanol. The membraneswere then allowed to air dry completely before adding scintillant.

The bottom of the plate was sealed with white backing tape, 30 μl/wellof Microscint 20 (Packard Instruments, catalog no. 6013621) was added.The top of the plate was sealed with clear sealing tape, and the platethen counted using the Packard TopCount with the appropriate settingsfor [³³P] with liquid scintillant.

Procedure for Phosphocellulose Filter Plate Assay:

Step 1:

The enzymatic reactions were performed as described in Step 1 of theStreptavidin Flash Plate Assay (above) utilizing KKGGRARTSSFAEPG(SEQ.ID.NO.: 16) as the substrate in place of biotin-GGRARTSSFAEPG.

Step 2:

The reaction was stopped by adding 20 μl of 0.75% H₃PO₄. 50 μl ofstopped reaction was transferred to the filter plate (UNIFILTER™,Whatman P81 Strong Cation Exchanger, White Polystyrene 96 Well Plates,Polyfiltronics, catalog no. 7700-3312) and the reaction incubated on thefilter for 1-2 minutes before applying vacuum.

The plate was then washed using a vacuum manifold as follows: 1) 9×200μl/well of 0.75% H₃PO₄; and 2) 2×200 μl/well of diH₂0. The bottom of theplate was sealed with white backing tape, then 30 μl/well of Microscint20 was added. The top of the plate was sealed with clear sealing tape,and the plate counted using the Packard TopCount with the appropriatesettings for [³³P] and liquid scintillant.

PKA Assay:

Each individual PKA assay consists of the following components:

-   A. 5×PKA assay buffer (200 mM Tris pH7.5, 100 mM MgCl₂, 5 mM    θ-mercaptoethanol, 0.5 mM EDTA)-   B. 50 μM stock of Kemptide (Sigma) diluted in water-   C. ³³P-ATP prepared by diluting 1.0 μl ³³P-ATP [10 mCi/ml] into 200    Tl of a 50 μM stock of unlabeled ATP-   D. 10 μl of a 70 nM stock of PKA catalytic subunit (UBI catalog    #14-114) diluted in 0.5 mg/mil BSA-   E. PKA/Kemptide working solution: equal volumes of 5×PKA assay    buffer, Kemptide solution and PKA catalytic subunit.

The reaction is assembled in a 96 deep-well assay plate. The inhibitoror vehicle (10 Tl) is added to 10 Tl of the ³³P-ATP solution. Thereaction is initiated by adding 30 Tl of the PKA/Kemptide workingsolution to each well. The reactions were mixed and incubated at roomtemperature for 20 min. The reactions were stopped by adding 50 Tl of100 mM EDTA and 100 mM sodium pyrophosphate and mixing.

The enzyme reaction product (phosphorylated Kemptide) was collected onp81 phosphocellulose 96 well filter plates (Millipore). To prepare theplate, each well of a p81 filter plate was filled with 75 mM phosphoricacid. The wells were emptied through the filter by applying a vacuum tothe bottom of the plate. Phosphoric acid (75 mM, 170 μl) was added toeach well. A 30 μl aliquot from each stopped PKA reaction was added tocorresponding wells on the filter plate containing the phosphoric acid.The peptide was trapped on the filter following the application of avacuum and the filters were washed 5 times with 75 mM phosphoric acid.After the final wash, the filters were allowed to air dry. Scintillationfluid (30 μl) was added to each well and the filters counted on aTopCount (Packard).

PKC Assay:

Each PKC assay consists of the following components:

-   A. 10×PKC co-activation buffer: 2.5 mM EGTA, 4 mM CaCl₂-   B. 5×PKC activation buffer: 1.6 mg/mil phosphatidylserine, 0.16    mg/ml diacylglycerol, 100 mM Tris pH 7.5, 50 mM MgCl₂, 5 mM    θ-mercaptoethanol-   C. ³³P-ATP prepared by diluting 1.0 μl ³³P-ATP [10 mCi/ml] into 100    μl of a 100 μM stock of unlabeled ATP-   D. Myelin basic protein (350 μg/ml, UBI) diluted in water-   E. PKC (50 ng/ml, UBI catalog #14-115) diluted into 0.5 mg/ml BSA-   F. PKC/Myelin Basic Protein working solution: Prepared by mixing 5    volumes each of PKC co-activation buffer and Myelin Basic protein    with 10 volumes each of PKC activation buffer and PKC.

The assays were assembled in 96 deep-well assay plates. Inhibitor orvehicle (10 Tl) was added to 5.0 ul of ³³P-ATP. Reactions were initiatedwith the addition of the PKC/Myelin Basic Protein working solution andmixing. Reactions were incubated at 30° C. for 20 min. The reactionswere stopped by adding 50 Tl of 100 mM EDTA and 100 mM sodiumpyrophosphate and mixing. Phosphorylated Mylein Basic Protein wascollected on PVDF membranes in 96 well filter plates and quantitated byscintillation counting.

Compounds of the instant invention described in Scheme 1 and Table 1above were tested in the assay described above and were found to haveIC₅₀ of ≦50 μM against one or more of Akt1, Akt2 and Akt3.

Example 5

Cell Based Assays to Determine Inhibition of Akt/PKB

Cells (for example LnCaP or a PTEN^((−/−))tumor cell line with activatedAkt/PKB) were plated in 100 mM dishes. When the cells were approximately70 to 80% confluent, the cells were refed with 5 mls of fresh media andthe test compound added in solution. Controls included untreated cells,vehicle treated cells and cells treated with either LY294002 (Sigma) orwortmanin (Sigma) at 20 μM or 200 nM, respectively. The cells wereincubated for 2, 4 or 6 hrs, and the media removed, The cells werewashed with PBS, scraped and transferred to a centrifuge tube. They werepelleted and washed again with PBS. Finally, the cell pellet wasresuspended in lysis buffer (20 mM Tris pH8, 140 mM NaCl, 2 mM EDTA, 1%Triton, 1 mM Na Pyrophosphate, 10 mM θ-Glycerol Phosphate, 10 mM NaF,0.5 mm NaVO₄, 1 μM Microsystine, and 1× Protease Inhibitor Cocktail),placed on ice for 15 minutes and gently vortexed to lyse the cells. Thelysate was spun in a Beckman tabletop ultra centrifuge at 100,000×g at4° C. for 20 min. The supernatant protein was quantitated by a standardBradford protocol (BioRad) and stored at −70° C. until needed.

Proteins were immunoprecipitated (IP) from cleared lysates as follows:For Akt1/PKBI, lysates are mixed with Santa Cruz sc-7126 (D-17) in NETN(100 mM NaCl, 20 mM Tris pH 8.0, 1 mM EDTA, 0.5% NP40) and Protein A/GAgarose (Santa Cruz sc-2003) was added. For Akt2/PKBθ, lysates weremixed in NETN with anti-Akt2 agarose (Upstate Biotechnology #16-174) andfor Akt3/PKBK, lysates were mixed in NETN with anti-Akt3 agarose(Upstate Biotechnology #16-175). The IPs were incubated overnight at 4°C., washed and separated by SDS-PAGE.

Western blots were used to analyze total Akt, pThr308 Akt1, pSer473Akt1, and corresponding phosphorylation sites on Akt2 and Akt3, anddownstream targets of Akt using specific antibodies (Cell SignalingTechnology): Anti-Total Akt (cat. no. 9272), Anti-Phopho Akt Serine 473(cat. no. 9271), and Anti-Phospho Akt Threonine 308 (cat. no. 9275).After incubating with the appropriate primary antibody diluted inPBS+0.5% non-fat dry milk (NFDM) at 4° C. overnight, blots were washed,incubated with Horseradish peroxidase (HRP)-tagged secondary antibody inPBS+0.5% NFDM for 1 hour at room temperature. Proteins were detectedwith ECL Reagents (Amersham/Pharmacia Biotech RPN2134).

Example 6

Heregulin Stimulated Akt Activation

MCF7 cells (a human breast cancer line that is PTEN^(+/+)) were platedat 1×10⁶ cells per 100 mM plate. When the cells were 70-80% confluent,they were refed with 5 ml of serum free media and incubated overnight.The following morning, compound was added and the cells were incubatedfor 1-2 hrs, after which time heregulin was added (to induce theactivation of Akt) for 30 minutes and the cells were analyzed asdescribed above.

Example 7

Inhibition of Tumor Growth

In vivo efficacy of an inhibitor of the growth of cancer cells may beconfirmed by several protocols well known in the art.

Human tumor cell lines which exhibit a deregulation of the PI3K pathway(such as LnCaP, PC3, C33a, OVCAR-3, MDA-MB468 or the like) are injectedsubcutaneously into the left flank of 6-10 week old female nude mice(Harlan) on day 0. The mice are randomly assigned to a vehicle, compoundor combination treatment group. Daily subcutaneous administration beginson day 1 and continues for the duration of the experiment.Alternatively, the inhibitor test compound may be administered by acontinuous infusion pump. Compound, compound combination or vehicle isdelivered in a total volume of 0.2 ml. Tumors are excised and weighedwhen all of the vehicle-treated animals exhibited lesions of 0.5-1.0 cmin diameter, typically 4 to 5.5 weeks after the cells were injected. Theaverage weight of the tumors in each treatment group for each cell lineis calculated.

1. A compound of the Formula E:

wherein: a is 0 or 1; b is 0 or 1; n is 0, 1 or 2: p is 0, 1 or 2; RingK is selected from:

Q is selected from: 2-azepinone, benzimidazolyl, benzimidazolonyl,2-diazapinone, imidazolyl, 2-imidazolidinone, indolyl, isoquinolinyl,morpholinyl, piperidyl, piperazinyl, pyridyl, pyrrolidinyl,2-piperidinone, 2-pyrimidinone, 2-pyrollidinone, quinolinyl, tetrazolyl,tetrahydrofuryl, tetrahydroisoquinolinyl, and thienyl, optionallysubstituted with one to three substituents selected from R^(6b); R¹ isselected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN,(C₁-C₆)alkoxy, O(C═O)(C₁-C₆)alkyl and N(R^(b))₂; R² is selected from:oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN, (C₁-C₆)alkoxy,O(C═O)(C₁-C₆)alkyl,(C₂-C₁₀)alkenyl and N(R^(b))₂, said alkenyloptionally substituted with oxo; R^(6b) is selected from:

optionally substituted with 1 to 3 substituents selected from oxo, OH,N(R^(b))₂, halogen and O_(a)(C₁-C₆)alkyl; and R^(b) is independentlyselected from H and (C₁-C₆)alkyl; or a pharmaceutically acceptable saltor a stereoisomer thereof.
 2. A compound of the Formula F:

wherein: a is 0 or 1; b is 0 or 1; n is 0, 1 or 2; p is 0, 1 or 2; RingK is selected from:

R¹ is selected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH,CN, (C₁-C₆)alkoxy, O(C═O)(C₁-C₆)alkyl and N(R^(b))₂; R² is selectedfrom: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH, CN,(C₁-C₆)alkoxy, O(C═O)(C₁-C₆)alkyl, (C₂-C₁₀)alkenyl and N(R^(b))₂, saidalkenyl optionally substituted with oxo; R³ and R⁴ are independently: Hand (C₁-C₆)alkyl; and R^(b) is independently selected from H and(C₁-C₆)alkyl; or a pharmaceutically acceptable salt or a stereoisomerthereof.
 3. A compound of the Formula G:

wherein: a is 0 or 1; b is 0 or 1; p is 0, 1 or 2; Ring K is selectedfrom:

R² is selected from: oxo, (C═O)_(a)O_(b)(C₁-C₁₀)alkyl, CO₂H, halo, OH,CN, (C₁-C₆)alkoxy, O(C═O)(C₁-C₆)alkyl, (C₂-C₁₀)alkenyl and N(R^(b))₂,said alkenyl optionally substituted with oxo; and R³ and R⁴ areindependently: H and (C₁-C₆)alkyl, wherein said alkyl is optionallysubstituted with up to three substituents selected from: OH and halo;and wherein R³ and ⁴ may be joined to form a (C₃-C₇)cycloalkyl; or apharmaceutically acceptable salt or a stereoisomer thereof.
 4. Acompound which is selected from:5-Methoxy-2(4-{[4-5-pyridin-2-yl-4H-1,2,4-triazol)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridine;2-(4-{[4-(5-Pyridin-2-yl-4H-1,2,4-4triazol-3-yl)piperidin-1-yl]methyl}phenyl)-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one;1-{4-[4-(3-pyrimidin-5-ylquinoxalin-2-yl)benzyl]cyclohexyl}-1,3-dihydro-2H-benimidazol-2-one;3-[3-(4-{[4-2-oxo-2,3-dihydro-1H-benimidazol-1-yl)cyclohexyl]methyl}phenyl)quinoxalin-2-yl]thiophene-2-carbaldehyde;1-(4-{4-[3(H-pyrazol-5-yl)quinoxalin-2-yl]benzyl}cyclohexyl)-1,3-dihydro-2H-benzimidazol-2-one;and2-[4-(1-amino-I-methylethyl)phenyl]-3-(2-thienyl)-1,6-naphthyridin-5(6H)-one;or a pharmaceutically acceptable salt or stereoisomer thereof.
 5. Acompound according to claim 1 of the Formula E, wherein; Q is selectedfrom:

which are optionally substituted with one substituent selected fromR^(6b); or a pharmaceutically acceptable salt or a storeoisomer thereof.